Mictal means man going their own way. But how can that be? It is already a movement and you follow a pattern that has been predictated to you. So obviously you're not going your own way. Alright guys, welcome back to the channel if you're new, my name is Bobby. Guys, today we're going to talk about being an alpha male. Yes, that terminology has been phrased by the red pill movement, by the Mictal movement. In the pickup artist industry people talk about becoming an alpha male. But even fitness influence us and generally people in their 20s talk about being an alpha male. Not always in their 20s to be honest. Some people don't get over it even in their 30s and 40s. But usually most of the time people in their 20s are interested in becoming an alpha male. And that is the topic of today. Can you become an alpha male? If you think about the terminology alpha male, you of course have to think about nature. Because this is where this terminology is derived from. We're talking about animalistic behavior. In the animal kingdom you can spot the alpha male. The male that is the most dominant, the male that gets the females procreates the most and is the protector of the pect. That is usually the alpha male. But if we think about nature we have to think about the communication as well. Because how is it communicated that the alpha male is indeed the alpha male? They cannot speak after all. Animals have no language. They have non-verbal communication. So if they have non-verbal communication, being an alpha is communicated non-verbally. First and foremost through fighting of course. If you see two animals fight over a female for example, that's how it is established who the alpha male is. But in some cases you see that some animals do not fight at all. It is the fight-or-flight mechanism after all and some animals simply flee. So there is no proving ground at all and those animals obviously are not the alpha male at all. Once the dominance is established the male gets to procreate and after that becomes the leader of the pect. Of course that will be challenged after a while. Then younger animals will challenge the alpha male. So hence being an alpha male in nature comes with a lot of responsibility, a lot of stress. Constant fight for survival, constant fight for females, constantly hunting, constantly protecting. It comes with a huge burden. And therefore you will observe that most animals actually don't want to fight. Most animals actually don't want to lead. You can see that if you have pets. For example your dog will constantly test you and will try to establish dominance because that is simply his nature, but ultimately will be very, very, very happy to see that you are still the peck leader, that you are still the alpha male. Because that means the dog can follow. If you look into humanity now you will observe that most people really love to follow. Especially in this day and age it couldn't be any more obvious of course most people are sheep and most people love to follow. Be it dietary advice, be it religious dogmas, be it politically, be it socially you name it. It is very, very hard for people to stand out and really go their own way. Mictal means men go in their own way. But how can that be? It is already a movement and you follow a pattern that has been predicated to you. So obviously you are not going your own way. It is so ridiculous, look at it. Men do not go their own way. Men follow examples, men follow other men. This is what you see in society throughout the ages. It is human nature, it is completely normal. We are social creatures after all and some people are simply not leaders. Most people are simply followers. So let's get back to the subject of being an alpha male. How can all those followers out of the sudden become leaders? The quick answer is of course they cannot. Being an alpha is something that is communicated non-verbally. It means it is something that is felt, something that is lived. The animal in nature that has been destined to be the alpha male, think about it, biologically destined to be the alpha male. It never searched on Google how to become an alpha male. It just happened that it won the fight. The genetics determined the outcome of that animal. The genetics became that animal's destiny.  Of course an animal cannot reflect but there is no doubt or uncertainty. The moment that there is any doubt or uncertainty that animal is not the alpha anymore.  Because that is something that you cannot overcome by following what other men have created for you. You can only follow your own true path and then maybe you can overcome your challenges and return to God ultimately, spoiler alert, but following concepts online will never resolve those underlying issues. Don't you see that those alpha movements create followers and this is the name of the game. It is a marketing trick. No other man can make you an alpha.    You will go to other man that you perceive having a higher value of you. You will learn from them how to become like them. You will never be true to your own nature and like that you will never be an alpha male and that already dictates that your true nature is a beta male if you want to use those terms. Those movements are created by little insecure men. When I was in my 20s I met a few pickup artists and I did pick up myself, met a few ladies. All of that is completely empty, completely hollow. I know that if you're 20 and you're listening to this, you won't get it yet and it's completely fine. You will have to do your own mistakes and I know that women are the most alluring thing when you're in your 20s being the alpha, etc, etc, etc. But ultimately it is so hollow. If you follow those guys that claim to be alphas, you will be just like them.  If you really want to overcome this deep hole inside of you, there is no other way than returning to God. If you return to God, you become a man of God. If you follow those guys, you stay a follower. Alright guys, and this is it for today's video. If you liked it, leave it a thumbs up. If you haven't subscribed already, guys, please do so and let me know in the comment section what you think about the whole alpha male topic. If you want to support this channel as well, guys. Links in the description box. Thank you so much for your ongoing support. Really my heart goes out to all the Patreons and everybody that supports via superchats, via streamlabs, offline donations, etc, etc, etc. Thank you so much. As always guys, may God bless you all, much love and peace.


Who says that inspiration can't come from the most interesting of places? Hey guys, so I said I'd do this review last week. Apologize for the lateness I'm gonna review the animated Wonder Woman movie. They came out in 2009 This is one of the best DC animated universe films They ever made this is when Bruce Tim was at the top of his game for producing Projects this was an origin story for Wonder Woman and this movie Well, let's just say that the new one took a lot of inspiration from this film This is an origin film it talks about the whole aspect of Steve Trevor and Diana her being introduced into the world of man Ares is the villain the whole Idea of not being able to trust man and everything like that. There are a few differences There is a little bit more of a focus on the Ares Diana is a little bit more kind of matter-of-factly with Her sort of assumption of what the world of men is and there's oh, yeah There's this giant battle in front of the White House of Demons and Gargoyles and Amazons This is another movie in the DC animated universe films that I kind of say that they should stick with PG-13 This film is violent. There's blood in it. Admittedly the PG-13 rating has gotten a lot more so Violent in recent films especially with the last one the Judas Contracts and Teen Titans film There was a lot more violence in that one and it was still PG-13. So anytime they ever say they're gonna do an R movie I don't understand. They are perfectly fine with PG-13 The film follows the same sort of arc as that of the newly released film It talks about Diana's origins it does specify much more detail how Ares has a connection with both Wonder Woman's mother as well as Wonder Woman herself and There is a much better Conflict between the two Admittedly the idea that she's going after who she thinks is Ares in the new film was an interesting idea This is a very straight-up film. She knows basically right off the bat what she's going into what she's after It is set in modern day There are elements from the Wonder Woman comics that are introduced into this film That don't work as well mainly the invisible jet They don't explain that at all in this film because I think it's near impossible to explain how an Amazon who lives on an island with Greek architecture and Society ideals as well as technology as a jet So that is kind of what I'm happy about the that isn't in the new films At least for the time being but aside from that I still really enjoy this film The voice acting is on par Nathan Phillion as Steve Trevor is pretty good His character is not as fleshed out as it was this by Chris Pine in the newest film, but he still does a funny job It's Nathan Phillion. You can't really go wrong with him in voice acting Carrie Russell brings a cool much more Adult attitude to Diana in this film. She's not as Innocent and as funny as she was in the live-action film We see the characters are much more of a leader much more of a stronger figure character that she's meant to be in the comics as Well as what she was in the animated Justice League TV show Ares is also as I said much more fleshed out in this Alfred Merlone as his voice actor is Absolutely great casting. He's a pretty cool villain The whole concept of his lore in the Greek mythology and into this movie is very well done I enjoyed pretty much all aspects of this film The there are a few things that are done better in the live-action the main thing being the romance and the relationship between Steve Trevor and Diana Much much better in the live-action film again This is a short animated film and more of the focus is going to be on Kind of introducing and setting up characters as well as then getting to the fight scenes and the fight scenes in this Are pretty cool, especially the climax I'll say the climax in this one is better than the one in the live-action Aside from the sacrifice of one character in the live-action. This one is just balls out.  You've got amazons. You've got flying horses. You have lightning bolts from gods There is everything you could think of that happens in this climax And what's so cool is how the beginning of the film reflects on how the climax ends And there's that mirror polarity and it's actually really cool to see as well as when I first watched it I actually had a oh I dropped my jaw and said holy shit because I didn't expect it coming I enjoy this film There are some aspects I think that could have had a little bit more of a weight to them a little bit more fleshed out like I said the romance and the sort of the characteristics between Steve Trevor and Diana but otherwise I feel that this is a good movie and There's a lot of aspects that the live-action film may have Incoherently barred from this film also the big thing is about this film is its comparison of Aries using this film as opposed to the live-action film and live-action film It's well used to an extent But it really kind of comes out a left field at the end not saying that I didn't mind that but I Feel that it's much better done in this film as by especially since we learn more about the character Also, there isn't that bullshit about join me Diana that was in the new one that I admit was really Unnecessary he's a god of war His whole point is to kill people not have people join aside. I don't know so in the end I still really enjoy the animated Wonder Woman film is one of the best ones in DC animated universe catalog It is a definite one that should be seen anyways My final rating for the animated Wonder Woman film is this six out of seven I enjoy this film more than the live-action one. I think it's a better film. Did I enjoy the live-action one? Yes, was it a Marvel movie basically in sort of its sense? Yes, but I still like both films But admittedly since the animated one came out first and a lot of the ideals from the live-action are borrowed from this film I have to agree with this one and side with it more. Anyways guys, that's all for me I hope you enjoyed this review. I wanted to talk about this movie for a long time. It is in my bucket list movies So anyways guys, that's all for me. See you guys next time


It's Diane. Ann, Ricky Karooch. Hey. Hey. Hey, what's up? Well, my dad tells me you're trying to get a deal under contract with me and I'm just wondering what property that is. Oh no. Oh, Dave. Oh, Dave Harbour. Yeah. Okay, have me confused. I was like, I don't think I'm doing anything with her. And I was like, what's her number? I'm going to call her right now. So guys, so already had so much, already had so many things happen this morning. For one, 430 ran, rode my bike 12 miles with Phil for an hour. Then went to the gym, worked out chest after a big breakfast and about to go live up in Zero to Diamond. By the way, so many people are crushing it with the Zero to Diamond coaching program. People are posting in the group and emailing me and all kinds of crazy stuff. Talking about how much like they're getting this listing or that listing. There was one guy that he said he'd been in the business seven months. He sold like two properties in seven months. He's been working my program for three weeks and he's written two offers. He has four listings. He's done all this business. He said his business has exploded since he started using my program. Yes, sir. Gosh, damn. Tell you what, it feels good getting more done before lunch. Most people do all month. What's up guys? Happy Monday, Monday fun day. Mondays are like Christmas to me. Like when I was like seven or eight years old, how excited you got the next day. Like Sunday is Christmas Eve. I go to bed. I can hardly sleep because like I get the jitters because of everything that's fixing to happen. Like I love going to battle. I love going to war. I've already hit my quota for the week. I think it's like not even nine o'clock. I've already put two properties under contract. I got a listing and I'm following up with so much stuff right now. It's crazy. It's insane. I love this new market we're in with lower transactions because all you have to do is adjust your game plan. You leave the other agents in the dust. So so many things happening. So many cool things. I'm going to be going live on YouTube Friday calling old expires at 3pm central. I knocked it back an hour. So it gave me two hours to call. So I'm really looking forward to that. I'm still working on the zero to diamond, the new online course for you guys. Really excited about that. You guys can go ahead and go there and check it out. I'm almost done with it. I'm actually restructuring the 90 day action plan right now. That's the last big piece of it. But I've already got all the courses and stuff done and lessons reconfigured and all that stuff. So you guys dig into it. Crush Monday, Crush Tuesday, Wednesday, Thursday, Friday, Saturday and Sunday. And reach out if you need anything at all. And by the way, I just told the blog that so many people are crushing it with the zero to diamond system. Scroll through the group and see all the people that are getting listings, finding buyers. And the business is going to another level because they're adopting this relationships over transactions philosophy.  And this losing deals is the best thing that could ever happen to you philosophy. So it looks like I might have froze up, but I don't care. Because it's Monday and I love Mondays and we'll talk to you guys soon. I have so many deals I can't even get to them all. It's insane. People don't even want to answer because they're not going to have to do a deal if they answer.


The Smart Healthcare Facilities Project saw the handover of 15 refurbished health facilities to St. Lucia in October 2022. The facilities have been upgraded to green climate-resilient structures. The project was implemented in a number of countries around the region, with funding provided by the UK government and the UK Foreign, Commonwealth and Development Office. A delegation from the UK government during a short visit to St. Lucia toured two of the 15 facilities. Malcolm Gehr, UK Development Director for the Caribbean Development Team, says the UK remains committed to providing continued assistance to the Caribbean in the area of climate change. The medical centres we've retrofitted have been with PAHO, the regional health organisation. And actually what we've done here is pilot a new model which didn't exist. So we've now designed something here with PAHO that we've proved. So far, fortunately, it hasn't been tested in St. Lucia, but in St. Vincent, for example, our health centre is there that we've retrofitted withstood the eruption and were opened very quickly after the eruption. The UK official expressed confidence in the outcome of the project, which he says can further benefit other vulnerable countries. The facilities received overhauls, inclusive of solar panels, rainwater collection systems and disability access. Women and Secretary in the Ministry of Health, Wellness and Elderly Affairs, Jenny Daniel, says a new maintenance structure within the ministry was necessary to ensure upkeep. So we recognise that we cannot do things in the same way that we have done in the past and in anticipation of the new demands that will be placed on us as a result of the smarting and the upgrading of these institutions, we have made the necessary adjustments to allow us to do so. Danielle says the ministry has already begun recording savings on utilities. She says the plan is to redirect these monies into ensuring the facilities are maintained in accordance with an established schedule. The UK funded the upgrade of 55 health facilities in the region, 15 of which are located in St. Lucia. At least three of the smart bills in St. Lucia have been awarded the gold standard. The Ministry of Health, Wellness and Elderly Affairs, I'm Sant Justin.


I'm George Church, and I'm a professor of genetics at Harvard Medical School. I'm also a member of the Broad Institute and the Vease Institute. I was born on MacDill Air Force Base in 28th of August, 1954, in Tampa, Florida. Yeah, there's one teacher that I've acknowledged several times, publicly, which is Creighton Bedford in ninth grade and eleventh grade, who's my math teacher, and he essentially let me off halfway through the year, both years, because I had some combination of my narcolepsy and the fact that I was, that I knew all the, I seemed to know the material, so he was big influence on me. Also, my photography teacher from tenth grade, John Snyder, was an amazing influence. And then, well, plenty of them in college, in graduate school, but notably Sun Ho Kim, who helped me transfer from my sophomore year in college to graduate school. Well, I graduated in my sophomore year after working with him for a year, and then tried graduate school at Duke with him, but then I flunked out, because I wasn't paying attention to courses I had already taken, kind of like high school, and I worked as a technician for a year with Sun Ho, so I was with him as an undergraduate graduate and technician, and then he said, you probably want a PhD, so I went to Harvard to work with Olly Gilbert. Yeah, so I worked on crystallography of the first folded nucleic acid, transfer RNA, which was the key to the genetic code, and wrote some software during that time that was in use for 30 years thereafter, but I also did some experiments, mostly software. Well, so some of the software I wrote as a crystallographer, I rewrote as a rotation student at Harvard, again, a crystallography lab, because they were the only labs that had scanners that could scan films, and all data was collected on films that find both X-ray and sequencing data, and then that software was, well, I was a consultant for BioRad for brief file, and it was completely rewritten for genome therapeutics in the late 80s, early 90s, and others used it for part of the earliest stages of the genome project. I think it's a mystery why Harvard would accept me after flanking out of Duke, usually it's the other way around, or nothing. I think it's because I was accepted at Harvard for application in the previous years, and also I had published five papers during the time I was flanking out. I had been on transfer RNA and also on a model for DNA protein interactions. And then Wally, he had come to Duke for a day at the invitation of graduate students. I spent almost the whole day with him because I thought of an excuse why I should be at every one of his meetings. I don't think it had much impression on him. He wasn't on the admissions committee or anything like that, but made a big impression on me. I was pretty, I was pretty sure even before he showed up that I wanted to work in his lab and do DNA sequencing, or possibly crystallography. But I decided on DNA sequencing. Well, yes, so while I was doing the crystallography of tRNA, there was one point we wanted to ask how general our crystal structure was, and did it apply to other tRNAs. So I typed in all the sequences available at the time, which is not something you would do today, and took about an afternoon to type in, almost all the sequences were tRNAs, and I typed them all in, including the modified bases. And I folded them up in the computer and I said, wow, this is really cool. I could get hundreds of 3D structures from one 3D structure and a lot of sequences. So sequencing must be a lot easier, and it's just as good. And so I knew somebody who was doing RNA sequencing at the time at Duke, a senior graduate student, and I decided that I wanted to do something much better than, much higher throughput than that. Maybe sequence every person's genome. And so I didn't really do the math right away, but it seemed like it was plausible. And so anyway, I went to Wally Slap, which at the time, they were just beginning to do DNA sequencing. And so at that time, sort of 50 base pairs was a big deal. So we had a ways to go. The main one that was in use, the one that I typed in all the sequences from, was RNA sequencing, where you would digest RNA, label it and digest it and run it down on paper chromatography or paper electrophoresis. And there was a lot of redactivity, typically, and a lot of high voltage for the paper electrophoresis and kerosene-like substance. And so this high voltage in paper and kerosene was in a room which had an automatically closing door to a bunch of CO2 jets to put out the fire, should that ever happen. And so then we started converting over to gel electrophoresis, due to work with Tom Maniatis and then later Wally Slap. It was in the mid-70s, 77. We were well in the era of gel electrophoresis. Well, so I think the average student would flounder around for a few months, even in the lab where you had protocols working. And last, we didn't have protocols working. They take maybe even longer. But then once you were up to speed, Greg Soutcliffe, the person who taught me sequencing, he was a six-year student when I was a first-year student, he knocked off 4,000 base pair, PBR322, that was a little bit of help from me in about a year. So 4,000 base pairs a year. And he was extraordinary. Fred Sanger was also extraordinary, but there weren't that many at the time. We almost never talked about cost other than when we ran out of acrylamide. And we used so much acrylamide.  It was like a $13,000 purchase order because we got it in quantity and high purity. And we used a ridiculously large amount of it until Fred Sanger published his paper on how to use thin gels with thin lanes and low percentage of acrylamide. And he clearly was being cost-conscious, but you wouldn't call it like a technology improvement. It was like three obvious cost-saving things that, by the way, also helped increase the read length. And when I said that 4,000 was a typical length, I mean, most people were satisfied with like one run up on a chest x-ray film, and so they might get 60 base pairs, and that would be their thesis at the time that I entered the lab. But in terms of communities, there really weren't that many. There were little communities centered around Wally Gilbert's lab and Fred Sanger's, and their trainees would slowly percolate out, and Wally's lab at least would send out these photocopied in a very colorful paper, green and pink paper that they would send out. I guess so people could easily find them on their bench among the clutter. Well, in Wally's lab, since it was one of the two centers, it was easy to find out what other people do. There was a little bit of traffic in between, very little. In fact, I think it might have been the first person in Wally's lab to do Sanger method, and a number of people said, you're going to be thrown out of the lab because that's the enemy, and I said, wow, we'll see. I don't think so. And he was fine with it, but that was kind of the limit of the flow of information. Well, I mean they were pretty independent, as far as I could tell. They both noticed the polycrylamide gel electrophoresis on the denaturing gels that Tom had developed, and they tried it out with different ways of doing end ladling, meaning, so if you just throw out where you'd get a ladder where each base was longer by one base per each, so the ladder could either be terminated by a chemical cleavage or by the polymerase falling off, but they resulted in the same sort of thing, but they were wildly different methods. And Gilbert's chemical sequencing took off early. It was a little easier to implement on double-stranded DNA, which was what most people had, SC40, PBR322. And Fred Sanger's was kind of restricted to single-stranded DNA, which PBR, sorry, FIX174 and G4 and M13 were a few examples. Anyway, so almost everybody had a double-stranded piece of DNA that they could label at one end, and so the chemistry took off for a while, but ultimately the didoxy sequencing once they had, once Joe Messing introduced single-stranded vectors that people could use that took off because of a slightly higher quality and no toxic chemicals. Still had PBR2 or S35. As I was leaving Duke, I was thinking about ways to change sequencing radically. And of course, this is not healthy for an incoming student because you just learned the protocols, but the ideas I had kind of in a vacuum because I was in transition from one lab to another were having to do multiplexing, how you can mix lots of things together, so the same volume would do multiple reactions. And I tried it a little bit during my rotation with Greg, and he said, no, no, just finish your project on this plasmid, and I did. I was fine, I was happy with that. And then I did one other project on RNA splicing, and finally I had a moment where I had to decide whether I was going to continue to do RNA splicing or really follow my dream of this technology development, and I decided that the RNA splicing I was doing would maybe impact three or four people worldwide while sequencing could affect more. And so I figured the ultimate multiplexing would be to do the whole genome in one tube. In other words, all the reactions could be done together. Running the gel, you could have the whole genome in one gel line, then the problem was like de-multiplexing it, sort of multiplexed it. And so the idea was to transfer to a flat surface. We tried a number, many different flat surfaces would work, and then probe it and image it, and probe it and image it. And that whole cycle of probing and imaging was the first inkling of next-gen sequencing. I mean, this was in 83 or something like that, published in 84. So people were excited about it. I mean, we had no idea what was coming with next-gen sequencing, but they were excited because you could do it without cloning or PCR, you could do it without implication, so you could get things like methylation and footprinting. So simultaneously, we gave away to do sequencing, methylation and protein footprinting, all from, essentially from nuclei of cells. So multiplexing part of it, I had had as a backup plan that if I couldn't sequence the whole genome in one tube in one lane, or then I would reduce the complexity of the mixture by mixing a bunch of plasmids that had your favorite inserts in them, and then you could do, let's say, 20 inserts, which is what I settled on later. But as it turned out, the whole genome sequencing did work, and it had many applications. In fact, somebody wrote a book about it, or a how-to manual, and then I said, well, the multiplexing might have certain advantages. Even though it's not the whole genome, it'll be more sensitive. I can now use non-reductive methods, because they were not very sensitive. But by reducing the genome size, I could increase the fraction of each target. And so that, I was multiplexing sequencing, which was the first paper I published at becoming a professor. In between, I had a short time as a postdoc at Biogen at UCSF, where I worked on stem cells, mammalian stem cells mostly. And then, but then that 1988 multiplex paper, along the way to that, I developed color metric sequencing, which was later used in, I think, all of the high schools in Seattle, thanks to Lee Hood, ironically using my color metric method rather than his fluorescent method, both of which were non-reactive, but I guess one was more expensive, it was early days. And then I developed chemiluminescent, and yeah, chemiluminescent, which was the main one that was used at Genome Therapeutics, and then with Peter Richterich coming into the lab was key in that. And that changed us from being the biggest reactivity user in Harvard to the lowest reactivity user, because we finally came up with a molecular biology tool, major one with non-reactive. And then fluorescence was the third method that we used with non-reactive. So the multiplexing was a step towards next-gen sequencing, and it was a step away from reactivity, and it was combined with automation immediately. Even the genomic sequencing, we really didn't automate it, even though I had developed the automation software back in 77, 78, it wasn't super popular. In fact, Greg, when he first heard that I was doing this, he says, what do you want to do that for? That's like the only fun thing of DNA sequencing is sitting down with your coffee and reading the gels. So I kind of put that on the side in 78, but it came back out in 88 when we did the multiplexing. Right, well, I heard that from myself. I was one of the first people to talk about it, but the first meeting where it was discussed that I know of was indeed the Alta 1984 meeting in Alta, Utah. I wouldn't call it organizational meeting. It was aimed at a different topic. It was kind of a hijacked Shanghai meeting where there was a very small number of like 10 scientists were invited. I was the youngest, and... It was an estimating mutation rates that might in some way be the consequence of atomic energy or other atomic bombs or so forth, or even non-atomic and other energy sources, anything that could cause it. And so there was a credential mandate to estimate this. We conclude in the first five minutes that it was not feasible at the moment, certainly, and the best we could do was maybe with a dollar base we could sequence human genomes, or actually a human genome as the way it was phrased, and that would eventually lead to another estimate of error induced by energy. But the point was we could do this, and we didn't know whether it was going to listen or not because we knew we weren't answering the mandate of the meeting, which also... It was sponsored not just by DOE, but also by the Office of Science and Technology. It was OST, I think at that time right at that OSTP. And it went back to DOE and went back to the Office of Science and Technology, and at DOE, they just started... They got excited about it, and they started writing checks. Mostly, internally, DOE had it. It had three small advantages. They were already up to speed on fluorescence-activated chromosome sorting, so they could make chromosome-specific libraries. They were good at bioinformatics because George Bell had done more or less what I had done except typed in a whole lot more sequences. And then they were also good at robotics because they had all kinds of robotics for handling radioactive substances and so forth. And then NIH had strengths as well, mostly in mapping, human genome mapping, and in model organisms. But at that point, nobody was talking about model organisms. It seemed like I was the only person at the first three meetings, one of which was not organizational, and you could argue the next two were. So it was the DOE, then the Santa Cruz, which was sort of independent, sort of university-run thing, and then DOE again at Santa Fe. And all of them were talking about the human genome. It was a human genome. It wasn't even a diploid human genome. It was just three billion base pairs. Pretty much all of them came to the conclusion of what dollar base pulled out of the air because I know the average student could not pull it off for a dollar base at that time. And certainly, it was completely neglecting the issues of repeats and scale and so forth. But they figured it was a dollar base. There was essentially no automation. There was little murmurings of automation in Japan. And Watson was like being very nationalistic. Jim took me aside at a wedding at Coltsbury Harbor. And the Japanese are going to just eat our lunch. They're going to take all of our economic wherewithal in genomics. If you don't do something about it, we just sequence E. coli. And I said, well, you know, great. I'd love to have them join. But anyway, they had Fuji was involved. And they were building sort of an automated process for making films, which were gels. So gels were automated, built more or less the way that film, photographic film was made. And so it seemed like a natural thing for them. And then they had a lot of other robotics manufacturers were interested. And I went over to Japan around that time. And it was quite impressive. It was like shock and awe of the day. But they dropped out. Fuji decided it was too flaky. It would hurt their reputation if they had bad quality of any sort. So they dropped out the robotics manufacturers. I think realized that having a robot do exactly what a human does is actually not cheaper than a human at that time. In fact, still to this day, if you want to do something with a robot, it's best to do it in a way that's quite different from the way the human does it. So when we got to next-gen sequencing, when we developed the first next-gen sequencing device, it didn't look like a robot at all. There was no robotics involved because everything was essentially again in one tube and spread out over a slide. And it was essentially microscopy. You had a microscopy with a lot of moving parts. So anyway, that was sort of 84, 84 or 86 when the first three meetings occurred. Right. So it seemed like it was going to be a real thing because in 87 they started giving out grants. I think I might have been the first grant, first genome project grant in 87. It was very modest. Everybody was joking about how I didn't know how to ask for a DOE grant because it was like $100,000 a year or something like that. And most DOE grants were in the many millions of dollars intramurally. But it seemed like a real thing then. And then NIH started getting excited about it. And then the whole biological community kind of said, no, we can't let this happen. We can't let NIH get involved. Don't worry about DOE because DOE was already this kind of this Byzantine, mostly intramural thing. And there wasn't really any way for a NIH researcher to get into the DOE. I mean, I came in straight out of the blue. I didn't even have a lab when I first started talking to them. And so when NIH started getting interested, then people freaked out and there was almost a letter every week to Science or Nature saying, this is a bad idea. I think it's been through a vote. It's probably been 99 to 1. But fortunately, Jim Lawson was involved and he was pretty charismatic and he went, not the word that everybody would use to describe Jim, but anyway, he went and talked to a lot of congressmen. And it became a separate line item, as I recall. There was also around that time, I think it was in 87, there was an NRC, N-A-S-N-R-C, to look into this. And Maynard and I were outside consultants. We weren't part of the committee. Maynard became part of the committee later. But that was, I think it was part of the process of figuring out whether you could do it or not. And eventually, NIH felt that they couldn't let Department of Energy run what could be the biggest and best biology project in decades. So they started working together and then it seemed like DOE was, I think, more technology oriented, but NIH, mostly their strengths was model organisms and mapping. It became a model organism mapping project, for better or for worse. And I think a lot of the mapping stuff was distracting. But they eventually got back to sequencing. Sequencing was in the first round of proposals. I was involved in three of the NIH and two of the DOEs. So in addition to mine, there was another one from Ray Gesslin's lab in Utah for the DOE. So we were the two DOEs that I can recall. And then NIH, oh, maybe four, at Jenny Mao, because collaborative research later becomes genome therapeutics. They were doing mycobacterium, lepre and tuberculosis. And then Wally Gilbert was doing mycoplasma, which is different from mycobacteria at Harvard. And then at Stanford, there was David Botstein and Ron Davis. And I've had many interactions with both of them, all of these people since then. And then the fourth one was Eric Lander. He was his first grant, my second grant. And these were all in the same stack. And they all got funded. And he did his own mouse. And it was mostly mapping. It was like four out of five specific games were mapping all the other three from NIH and two for DOE. We were all about sequencing. And then he had one sequencing section, which was written by Lenny Garanti and I. And even it was only partially sequencing. It was mostly CDNAs in yeast or something like that. Just a little bit of sequencing in it. But as soon as we got the grant, that started to take over, at least all the mapping they would do by sequencing if they could. And then it grew from mouse to human. So I would work with them because of proximity, but I would work with the other teams because I was interested in sequencing. And it ended up, most of the sequencing was done with my methodology. Well, there were all five of the six published using the multiplex sequencing at some point. But the one that did the most with it was collaborative research with chemo-therapeutics. They actually sequenced several whole genomes with it. So I knew Eric well before that. He was in the Harvard Biolabs. When I was a graduate student, I was kind of a visiting professor from the business school, I think, and he was hanging out with Peter Churbis and Bill Gelbart. Bill later went on to have a major role in databases, even though at the time he was a fly geneticist with very little interest in computers. But anyway, Eric would hang out with Bill and learn genetics. And then I went off and did my postdoc and didn't have much contact with him again until I came back as assistant professor. And then we started talking again. Around that time, he was doing a lot of work. He had done a series of rotations, you know, even though he was a lecturer at the Harvard Business School. He had worked not only with Bill and Peter, but also with Bob Horvitz and with David Botstein. I think one with David was the most successful of his rotations, where he did a lot of the math behind ideas that David Botstein had had for, I think, for years. And they together were able to implement these things in either math or software, or both. And so he was well-known already, even though he had not done any experiments. You may have still not done any experiments as far as I know. But anyway, so when we developed this center together, I think he was one of the biggest in that most of the other ones were picking a human chromosome. And I think it was smart to pick a whole genome, because as it turned out, a whole genome approach was a better approach than the chromosome approach. And in the end, the human genome was done by a whole genome. In fact, in a way, we weren't aggressive enough. It should have been a whole genome shotgun from the beginning, which was what I was advocating all the way through. But at that time, around late 80s and early 90s, almost every genome media you would go to, all the technologists, I mean, there were very few technologists, but all the people who called us the technologists were aiming for one X coverage. That was like the Holy Grail was one X coverage. And I just would shake my head and say, are you kidding me? The Holy Grail should be bringing the cost down so you can do any X coverage you want to. But that was just very unpopular sentiment at the time. Even though Fred Sanger had already developed shotgun sequencing, but to some extent, Fred was not a major component of these conversations. He was on his way to retirement, and I think most people considered him like a freak of nature that could do things that nobody else could do and just who knows. And he was much more of a technician than a leader. I mean, he certainly had leadership skills in the sense of setting an example, but he didn't have a big lab. He had typically one technician in the lab and an occasional visitor his whole career, even though he got two prizes. But anyway, when he would come to visit our lab, he would come sit in the room that I worked in and he would spend all his time talking to the technician in the room. He'd be talking about how much Tmed they used and what percentage gels and all this sort of stuff. He was very focused. So, anyway, so Fred's shotgun sequencing did not have a big impact at the beginning. And one by one, Easties Labs had to learn the value of 7X coverage or more. Now, all the organs were great. I was super... In fact, I think I was the first one at all the first three meetings. I said, no, we need genome sequence comparisons. Comparisons means you have to have something to compare to. You'd have two genomes, some will be closer to you, some won't. And I said, and we might as well start with small ones so that we get some payoff early on rather than spending 15 years and then hopefully having a payoff at the end. Let's get a bunch. That was not popular either. Most of the things I was proposing were not super popular. But what did happen is it was almost... In order to deal with the critics between 87 and 90, between the DOE and NIH starting, it was very politically adept to bring in my organisms. That's what helped change David Bonstein from a critic into a supporter of the sequencing. He was always high on genomics and genetics, but to specifically support the genome project, it helped to have yeast as part of the game. And then worms were obvious because Sulston and Waterston had already done a lot of mapping. I mean, they basically had all the clones in hand to go. And that's also what influenced the enthusiasm for mapping is because it worked so well in C. elegans. It didn't work so well in humans. And so those were obvious small organisms. Flies came in surprisingly late in the game. And then bacteria got in as an organism. It was very funny that we called it the bacterium. And it meant a bunch of bacteria, mycobacteria, E. coli, homophilus, mycoplasma from both Wally's lab and Craig's and so forth. So they were all lumped in together as if it was one organism Yeah, he was very significant, I think. So when that 1990 grants went in, where I worked with Jenny Mao and Ron Davis, David Boston were together and then Wally and Eric, the Boston Davis grant had in it was a beautiful grant. I think it was the best of all the grants that I saw. It got the worst score. I mean, it was essentially rejected, but eventually it was overridden. But it projected pretty accurately in a detailed manner all of functional genomics. It was all in that grant. You know, just all this beautiful biology and technology and with yeast is the perfect way to test it out. It was at least a U-curio. It was better than all these bacterial genomes. It was just a beautiful grant and it just got trashed. But Maynard and I were there for the site visit and we could tell from the very beginning site visit that they had just came in loaded with loaded shotguns ready to take us out. And Ron, so at the time there were two ways of doing microarrays. One was Pat Brown's, it was Alpha Metrics, and Ron had bet on the Alpha Metrics, which I think in the end was the correct bet. In fact, totally correct bet was not a raise at all for sequencing or for RNA analysis. It's still useful for SNP analysis. But anyway, Ron just went on to do all kinds of innovative technology and the legacy of that center was Ron's Technology Center which is still in existence today. And just so many interesting technologies have come out of there. Well, like I said, I mean, he was big on nationalism. He was persuasive on getting Congress to vote for it, for getting NIH up to speed. At that time it was not a institute, it was a center. And I think he led to something that was a serious enough effort that it had to become an institute. And I think he recognized the value of the genome from very early on. I don't remember him being one of the doubters that had to be convinced. Yeah, he seemed pretty compelled from the beginning. Other than that, I don't really remember much about his role. I mean, Coltsbury Harbor was one of the places where annual meetings would occur. And Coltsbury Harbor was also some of the grantees. But I don't think that had anything to do with why he did. I think he felt it was the next big thing to do. Coltsbury Harbor had a good meeting structure already for courses and meetings, mainly during the summertime. Including meetings that he had gone to when he was a postdoctoral fellow. And he announced some of his work on the DNA structure in the early days of biology. So he had a warm feeling for Coltsbury Harbor and became the head many years later. And it had an infrastructure for having exciting meetings, relaxed meetings. So I think that that and Santa Fe became the two main meetings. There were also meetings held at Hilton Head, called GSAC. You know, sequencing analysis conference. They tended to be a little more focused on sequencing a little bit earlier while the other meetings were mapping and sequencing. But I went to all those meetings for a while until they got into heavy, heavy production sequencing. I got lost interest and went to the other things. Yeah, very, very rarely, almost not at all. I mean, it was not an NIH grantee, right? So it was a DOE grantee. It still am since 1987. I never got an NIH grant of my own until 2004. Right, the SAICS. The SAICS grant, yeah. I think that might have been part of it. I mean, I, in a certain sense, had one through my collaborations with collaborative research and Stanford and MIT and Wallis at Harvard. So in a certain sense, I had four. But I didn't, I got zero dollars out of any of those four. I mean, to my lab, interestingly. So Eric got 19 million. I got zero from that first grant. So it shows how good a businessman I was. But I was very dedicated and I worked hard on any one of those who wanted me to work on it. But it didn't land me as an obvious advisor to NIH. So I was an advisor to the NRC, the NASNRC in 87. But I didn't really get that heavily involved in the NIH efforts until we got to SAICS. And also, when the $1,000 genome launched roughly around there during Zerhouni's era, I think Francis got the bug of the one called Grand Challenges, what were they called? Roadmap. Right, yeah. Anyway, there were like 11 different possible roadmaps. And I showed up, I did advise on that one. Re-sequencing the biome. No, no, that was one of them. But there was another one on technology. And I was, it's one of the few times I was pretty emphatic. Actually I was pretty emphatic all the way through the project. I mean, my meek version of emphatic, which is they should invest more in technology. Because it was my gut feeling that the return on investment in technology would be bigger than the investment of the sequence itself. Both of which were significant. But I think that once they got serious about technology, the price plummeted by three million fold. And almost none of that price plummeting was due to, was traceable to anything that happened during the genome project. In other words, there were a lot of things, a lot of technology development during the genome project, kind of minor incremental things. But it all just went out the window as soon as NextGen came in. Because the only thing that we really used from the old days was shotgun sequencing, which is we used big time in NextGen. But shotgun sequencing predated the genome project. It was, in effect, was mostly ignored in the first four years of the genome project. So anyway, I think that there should have been more technology development from day one. And finally that was listened to around 2000, I don't know, two or three when they started, they started thinking about the project, the thousand dollars. And even the thousand dollars was too radical. They had to couch it in terms of a hundred thousand and a thousand dollar project. So I probably liked Craig more than most people throughout the years. We would keep kind of tackling the same problems. I think mostly independently. So we both were attracted to small genomes early on. And arguably the collaborative research did the first small genome, which is a helicobacter. But Craig and Ham did the first really peer-reviewed one, which was homophilus. It was in 94 and 95. So that was one example. We also both got the photosynthesis bug about the same time as part of the DOE. We both also did Ham, including Ham Smith as part of this, because he was essentially second in command to Craig and did a lot of the real technology side of things. Ham was the one that made all the shotgun libraries. And he was the one that proposed synthesizing a genome. And he got DOE funding for that. Even though they hadn't put out an RFA for it. And so then I did the same thing about the same time. And the personal genome project was very similar to Craig doing himself. The difference was I got the approval and he didn't. But there were same kind of ideas that you could have an identified individual. He started out not identifying himself, but then later did. We started out identifying ourselves. Anyway, there were many times where we would do similar things. The biggest difference that he's acknowledged publicly is that I was much more into technology development. He would say in meetings, George will develop technology and we'll use it. Which was very, I thought very gracious of him. And I tried to return the compliment as much as I could. And so, you know, and he was, he was an early adopter of everything. And I think that really distinguished him early. I think that really was the first thing was in 87, I think, he had an NIH lab full of ABI equipment. He was a protein chemist. And so most of the ABI equipment was protein. And he had a big budget, but very little room for people. I mean, typical NIH budget, you could get equipment, but not people. And so he had to have like really automated equipment. So when they came out with the DNA sequencer, he didn't really feel that he absolutely needed one. But he felt, why not? It's another ABI machine. Let's get it. And he got one of the first ones. He even got one before Lee Hood did, even though Lee's lab had developed it. They didn't have the budget to buy one, as I understand it. But Craig did. He got one. He was a pretty good biochemistry know-how. And so they optimized the protocols a little bit. And he did something very clever. So he's still at NIH. He ordered the CDNA clones from Clone Tech. So CDNAs was a good choice, first of all, because it got rid of all the junk DNA problems. So 100 times smaller problem in principle. He then sent the CDNA clones from Clone Tech to Collaborative Research, which is where I had my center. And they would take it in laundry. They would do contract work for other people. So they made plasma preps for them. They would send the plasma preps to NIH, where ABI technician, together with one of his technicians, would run it on the ABI machine. So essentially he was using a third company to do the sequencing. And then he would run it without, I mean he wouldn't, he would run it without annotation, without proofreading, straight into NCBI. And so the whole thing was like almost a paper project. It's like Clone Tech, Collaborative Research, ABI in CBI. And to some extent just had to like make sure, just do little quality checks that everything's going okay. And I thought that was brilliant. But a lot of people got angry, that was one of the first things they got angry about. Before they even got angry about the patents, which wasn't his fault, they got angry about the quality. Because a lot of the reads were going in and they weren't even human. They were unreadable. They were, or the good ones had a 3% error rate, right? And so a lot of people said, ah, shouldn't even be in the database.  And so the people who had the prepared mind knew how to use it. So anyway, then the NIH did what a lot of public and private institutions did, is they started patenting this because it seemed like it was a good thing. And I'm not sure whether Craig, I don't think Craig initiated that. I think he went along with it. And then that got press. I was like, oh, this is horrible. We're patenting it, but you know, even though it was these low quality CD&I reads, which didn't have an application. You really need to have an, it had to be not only, not obvious, but useful. And a CD&I read was not, I mean, other than what people were doing, which was searching databases with it. And so that got, the interesting aspect of people getting upset about things is user results in the opposite of what they want, right? The more you raise the alarm in public, the more likely it's going to get more money. So the three examples that I was very close to was the common DNA. That attracted so much attention that it basically sparked investors to invest in Genotech and Biogen and Cetus and Amgen and so forth. And just, boom, out of nowhere, they got excited about Biotech. And then the CD&I, the Backel, and then the stem cell. For eight years, NIH wasn't funding stem cells. So California raised $3 billion. I mean, I'm not sure there would have been $3 billion spent in the whole United States on stem cells if there hadn't been this ruckus. So anyway, so finishing off, you know, where I feel Craig's role in all this was, that was the start. I mean, he was really not well known prior to that. But then he formed these related non-profit, for-profit, which was Tiger and HGS. And that was a brilliant business deal right off the bat because the HGS funded Tiger. All Tiger had to do was give them CD&I sequences. Eventually, Craig got tired of that relationship and he bought back Tiger so it could be an independent institute by that time getting lots of grants. And then, I think, then he had this race for the genome, which I didn't think was, you know, I mean, it was just using available technology. He had an inside track with ABI, which was a good thing, obviously. ABI's got a better chance of winning, fighting with its customers because it has unfair. So that was brilliant, too, business strategy. And then after the genome project was over, then he got disinterested in human genetic or sequencing pretty much for a while. He went off into synthetic biology using sequencing as a leverage to get people interested in his synthetic interests. He would sequence metagenomes and things like that. But anyway, so in terms of the human genome, I'm not sure it probably was a good thing to get kind of forced shotgun down everybody's throat. I think he did that. I tried to do that, but I was completely ineffective. He was very effective. And there was a period of time where people were doubtful that you could do holding a shotgun on human a mouse. In fact, I was part of a paper on how to do that, a computational paper. Gene Myers and James Weber from Marshfield did a paper. I was a co-author in that, but it took my name off at the last minute. I just don't know why I regret it. But I believed you could do shotgun on any size genome. And anyway, so he forced people to agree. And even after he did the human a mouse genome, there were still people arguing that it was not feasible or, you know, there was a P&S paper written by Eric and I think it was Bob and John where they kind of like critiqued the whole process. And I said, why don't you and your critique address the mouse genome? Because there really wasn't much mouse mapping, scaffolding at the time you did that. And that kind of showed that you could do it. And they just didn't want to have any part of that. And at the time, the reason they asked, or the reason Eric asked me was that at the time I was the only person who had both the venture and the, I guess it was the Santa Cruz Golden Path or Golden Gate, I can't remember, version in my lab. I mean, the two sides were not sharing sequences, but they were both sharing one with me. And so nature asked me to compare the two. And so they wanted to know whether I felt that the one was a derivative of the other. And in my opinion, it wasn't because when I went and got the Santa Cruz genome, it was actually in shambles. And this is not, I think it's documented in our paper, but not very aggressively. So what we did was we went to NCBI where they had quietly put in an FTP site with no fanfare whatsoever an alternative assembly of the human genome. This is in 2001 where there was a science paper and a bunch of nature papers. And we looked at the two and we said, oh man, the NCBI wants so much higher quality. We don't want the public to look bad. We want to have to put the best foot forward. We'll just kind of just call it the public genome. And I don't know if you saw the movie Contact, but it's a Carl Sagan thing. And they build this machine that allows interdimensional travel and some activists destroy it. But there's a second machine that's built in some, just like Rainy Island in Japan. Anyway, that's what this was like. This was like the second genome. And the reason Santa Cruz was so messed up, well, first of all, they rushed it with a small staff, basically one guy who was a hero who was painted as a hero. But also there were some bits that were flipped that made the assembly hard that were NCBI bits, but they had misinterpreted what they meant. Anyway, we compared the two and they looked pretty good. They were very similar but clearly independent in my opinion. So anyway, the critics didn't want to hear that and I didn't really care. And I think the answer is now, everything's done by shotgun. De Novo sequencing and resequencing. It's hard to say because I think the main innovation there was the 3700 instrument. And it's possible that would have galvanized everybody to go forward. It probably would have taken a little bit longer, but when you consider the goal was a polished genome, I think we might have gotten a polished genome faster, might not have gotten a draft slower. So in other words, rather than a draft in 2001 and a polished one in 2004, we might have gotten a polished one in 2003, actually faster rather than slower. And the polished one got almost no attention whatsoever. And the other thing that might have happened is there wouldn't have been such a panic. I think we're just kind of warming up the idea of technology development. If there hadn't been a panic, then it might have been fewer 3700s bought and more alternative sought. And we might have tried to finish the genome rather than produce a draft. I think all that were possibly unintended consequences, negative unintended consequences of a race. And part of it I think was stimulated by there was a round of review where there were five centers, which was one in Cambridge, Massachusetts, one in St. Louis, one in Baylor, Texas, one at Collaborative Research and one at Tiger. And they dropped the two that were not classical academic laboratories. So Tiger was non-profit, but it clearly was part of a for-profit institution and CRI was definitely for profit. They dropped those two and I thought that could have resulted in more innovation, or at least a different way of thinking, and also that stimulated Craig to go up and do his own thing. So I think that decision of dropping it from five centers to three was a poor management decision, maybe with hindsight. But I think if there hadn't been a race, there might have been more, the one option would be who's got the better genome, rather than who's got the first genome.  I hope to fix that soon, but no one's really encouraged, nobody's particularly encouraging of that, right? So you have to kind of do it on your own. Yeah, I think we should remind ourselves that it's not done yet. And it's some of the most interesting parts of the genome, I think. The centromeres could easily be involved in aneuploidies, in abortions, low birth weight, cancer, all that has to do with segregation of chromosomes, and that's where the centromeres are all about. So I think it can be done. In fact, I think the method that will get the centromeres and the other gaps will be the method that will displace all the other methods, just like the next end did. I knew Francis from his work on jumping, genome clone jumping, and also from 65 roses, and a number of things he had done before he came. I didn't expect that he was necessarily going to be a fantastic manager. You know, it just seemed like he was a regular postdoc, like I was. But I know Eric was quite in favor of it, and I think convinced Francis to try for the position. And it turned out I think he was really the leader that we needed. He was sufficiently into the science that he knew what was right and wrong and could help steer things, but wasn't so micromanaging that he would interfere with things. I think overall the NIH staff played a bigger role in those grants than almost any NIH staff in the history of NIH would be my guess. I could be wrong, but in terms of extramural grants, they were often co-authors on papers. I mean, that's a pretty deep involvement. And Francis kept a very active research of his own, which I think was great. Now whether, I don't think, you know, I don't know what it would have been like with somebody else. Clearly, you know, I think Jim might have been the perfect person to get Congress to vote for it, because he just had more credentials than Francis. I mean, it was a Nobel Prize winner at a very young age, and head of Cold Spring Harbor and so forth, he had some credentials. If Francis had come in, at the time Jim had come in, that probably wouldn't have been so good, even though it would have been a less rocky, there wouldn't have been the rocky exit. But I think Francis was the right one for that point onward. So I've already mentioned, I think some of the false starts were the mapping. Mapping turned out to be less interesting than it seemed at the time and consumed almost all of our resources for the first five years. So even though we ended up ahead of schedule, we would have been maybe a little bit more. And then the race, I consider a false start, because it discourages us from going for quality and it discourages us from technology development. I think the next-gen sequence, so who was involved? Sydney Brenner developed this MPSS method, which used beads, and I think the ultimate answer was in flat surfaces, not beads, but he was clearly a pioneer and around 1994 he published or patented MPSS, formed a company called Lynx. Lynx then licensed some of my technology, some of the multiplex tagging and strategy and I consulted for them briefly and then they merged with Selexa and Illumina. And at the time, what was interesting is when those three companies merged, they didn't use any of their technologies. So Illumina was basically a bead snip company. Lynx was a bead, subsequently by the Legation Company, or cleavage in Legation. And then Selexa was a single molecule sequencing company in its original instantiation. They basically threw out all those, maybe kept a little bit of the microscopy idea behind Lynx, and then in licensed some chemistry and amplification methods and then just started running with it. And there's still some dispute as to who invented the stuff, they had licensed it from somebody maybe other than the inventor. So Jing Wei Zhu was clearly a pioneer in all of the next gen in developing good reversible terminators for, so getting peer reviewed articles. I think we have to remember the peer review, not get so entangled with patents on reversible terminators for fluorescence sequencing and later reversible terminators for nanopore sequencing, which I think is still in the works, but it is promising. So the whole idea of sequencing by synthesis has been greatly helped by having those terminators. What else? For quality, most of the quality came in with half a typing, oddly enough. And because in a way it could have gone down with short reads, because there's mostly quality, most errors have to do with replacements. So if you try to put something that's mildly repetitive onto a scaffold even, it doesn't know where to go. But then half a typing, and I think the first really good half a typing was Complete Genomics in 2012 where they figured out how to break five to ten cells up into fractional genomes into 384 well plates. And then each of those would be read, which was kind of a mixture of 100 kb fragments. And it was as if you had done BACs, which by the way was also an innovative thing. Early on in the genome project was back sequencing, back in cloning. Yaks were a bit of a distraction, but BACs were really the real thing. We had an excellent BAC library from Peter DeYoung very early on, and for some reason it was sidelined, didn't use it. I think it was because it was Peter DeYoung's genome, and so he was a known individual, but they could have gotten an IRB approval for that. Instead they tried to make a diverse set of BAC libraries which more or less failed and ended up with a non-diverse single person. Again, just a de-identified single person, which I don't think is necessarily a plus. But anyway, the BACs were good. The not having the clone of the BACs was even better because then there were some cloning artifacts you would get and some sequences that weren't easy to go into BACs. If you just fragment up the genome and put it into a in vitro amplification, you ended up retaining more of the genome. That was 2012. The Complete Genomics had published a previous paper in 2009, which was really the first, I think, truly inexpensive genome. They had consumables, meaning reagents and supplies and equipment amortization, on the order of $1,500 per person back in 2009. It ranged up to $4,000, but it was in that. It wasn't quite the $1,000 genome. Who else was... I think that's it for innovations and false starts off the top of my head. I think there's relatively little logic because there are relatively few labs that do it. In academic labs, it's much more tempting to be an early adopter. You get labeled as a technologist if you're an early adopter. In industrial labs, there's also a great emphasis on incremental growth and in licensing things that are invented elsewhere. Typically, elsewhere will be some kind of hole-in-the-wall technology group that developed one particularly cool thing. It might be like Stan Tabor and Charles Richeson developed Sequinace. That made its way into the genome project for a while. Matthews and others developed capillary electrophoresis and that got brought into EBI. There weren't labs that were really full-time doing technology development. They might be doing mostly analytic chemistry or mostly biochemistry, and they'd have an insight and then get in the lenses. There are a few labs that were full-time technology. Obviously, Ron Davis' lab, Lee Hood's and mine were arguably three of those labs. It was hard to be academic or industry. You had to be at the interface a lot, which is hard. In terms of patterns, so patterns within that, the general pattern, most technologies get displaced, sometimes without a trace other than the history. It might last about a decade. Decade is a good length of time for technology to last. Our multiplex sequencing lasted about 13 years. Sanger's radioactive sequencing lasted a similar period of time, but depending on how you count the fresh start with the EBI fluorescence sequencing, it lasted a bit longer, but it all depends on where you... I would consider it almost a totally new method, because you're not taking the plates apart and slapping an X-ray film on it. There's just a whole lot of differences. You're running everything in one lane, other than four lanes. The only thing that's so different, the only thing that I have in common, was even the didioxies weren't the same. They were the determinators and other innovations. It takes about five years to go from a concept and maybe a preliminary paper to an instrument. That's a rule of thumb. Then that whole technology will last about a decade, and another one will come in. Sometimes it'll build on top of it's like the EBI did. Other times it'll completely displace it like next-gen sequencing. There's not almost a trace left of the electrophoretic era. Probably the next one might be nanopores. That doesn't quite fit the pattern perfectly in the sense it has taken a whole lot more than five years to get from a concept, sort of like 88 concept, to barely working in 2016. The first human genome was essentially sequenced with nanopore in 2016. The first bacterial maybe a year before that. That's kind of the gold standard now, is can you sequence a human genome at fairly high accuracy? That took from 88 to 2016 to arrive. I don't know how long it'll last, but I think it has the highest probability of displacing the current big iron, because one chip you could probably fit, well, you could already fit 8 million nanopore sequencers, and that chip is cheap and reusable, so you could imagine, and that could scale up to billions the way I and Torrent did. So anyway, those are the patterns that I can see. Well, part of it was single molecules harder than multi-molecule, so it was pretty noisy, and it really required a complete redo. I mean, both of the way people thought about electrophysiology, so when I did the first patent on nanopores, I was thinking patch clamp, and patch clamp doesn't scale at all, so the idea of changing, I mean, just like next-gen sequencing required a rethink from electrophoresis to flat on a glass slide, we needed to move the patch clamp from this sort of artistically crafted pore to something where you could have millions of them on a flat surface. In all cases, we were trying to ape, mimic silicon valleys or micro fabrication in electronics, where you have these super flat layers, and you put, and you use some combination of photolithography and self-assembly to put down the nanopores. Anyway, even with hindsight, that probably was going to take a long time. The other question is... It probably isn't that big a loss because right now, even with all the computer technology we have right now, the great limiting point for the genia nanopores is dealing with the data because you're getting terabytes of data from a little chip in short periods of time, and so we need to, and if we had had those terabytes of data back in the 80s, it wouldn't even be conceivable to deal with it, right? Now, at least we could kind of trim off. We can start building a field program with Gator-A's and GPU's right onto the chip and do a lot of data processing on the chip, but a lot of it's... If you do it really cleverly, it's incompressible data. I mean, the short answer is I think the $1,000 genome was distinctly better than everything else for technology development. And I was not a big fan of chips even though I was one of the first adopters of chips for a few demonstration articles. I was never... To me, I was just playing whether to see if there was something there. And the main thing that I got out of chips was stripping the DNA off the chip. So I used it in a very perverse way is rather than having them nicely ordered, stripped them off and started building big pieces of DNA out of them, or using them for making libraries. And to this day, that's the only thing I use chips for is synthetic biology, not for analytic, which was the point of most of the NHGRI efforts. I think NHGRI didn't get onto functional genomics early enough. I think the ENCODE project once it got started was terrific. In a way, it encouraged technology development. Again, not as aggressively as the $1,000 genome. I think the SEGS grant program was the best of the best. It encouraged out-of-the-box thinking. It encouraged interdisciplinary, multi-teams, not just multi-collaborators or multiple people, but actually multiple teams putting together innovative things. So it encouraged innovation, which is something you don't see often enough. And as it happens, the SEGS grant happened to fund me for next-gen sequencing before the $1,000 genome grant program started. And so when it started, I didn't apply for the $1,000 genome grants because I already had the grant and I didn't want to do double-dipping. But I thought the two of those were complementary. And if I had to choose, I would, at that point, pick the SEGS. I mean, I'd already picked the SEGS before I knew the $1,000 genome project grant was going. But I still would have picked it because I think it was more intellectually exciting. I mean, I went to almost all of the $1,000 genome seek-tech-dev meetings. In fact, I was on the grant-review committee. I was chairing the grant-review committee for a few years. And I loved them because they were full of physicists and engineers. But the SEGS was full of innovation and biology. So they were very complementary. They had two real jewels in the crown. Not just of NHGRI, but of all of NIH. The only thing that comes close to those two programs, in my opinion, are the transformative awards and the pioneer awards. Because they allow you to do things that would not normally get funded in a particular institute. Most of the really cool things that people celebrate or at least I celebrate are these interdisciplinary things that are cut across all the NIH institutes. Well, like I said, they had it sitting on their plate with the Boston Davis grant. And the fact that that grant got rejected by peer-review there in square, it's just the wrong peer-review. And then it got re-instantiated but greatly reduced in budget. I think that was the first sign. And then when I was on the grant reviews for some of the genome, early genome project grants, we were given strict instructions, no biology. And I think that was partly to develop a distinct portfolio for the NHGRI relative to the other NIH institutes and later they started having like joint grants. I thought that was brilliant. It allowed more biology to come in and it also effectively created bigger fund for NHGRI related projects. But I think that was partly the zone of inhibition from the other institutes that prevented NHGRI from doing functional genomics very early. And there was there were always clever grantees to figure out how to sneak it in. But and it is certainly quite healthy now for many years but took a while. So, nanopores in terms of technology, in terms of biology and technology, I would say in C2 sequencing and synthetic biology. I remember we were a SEGS meeting at Stanford was one of the first SEGS meetings and Francis was there and there was a Q&A period where people in the audience asked Francis questions and I think Roger Brent asked the question of would NHGRI consider funding synthetic biology? And Francis gave a pretty quick answer which was no. If I raised my hand and I said what if we had a program where you would do make targeted mutations to see what variance of unknown significance what their physiological effects would be. So that would be sort of synthetic biology but it would be testing the hypotheses that are flowing out of the genome project. And he gave a quick answer which was yes. So it's a really kind of way it was framed and that was the basis essentially for our second SEGS. So the first one developed next-gen sequencing starting in 2004 and the second one was on testing hypotheses using genome engineering which we're proposing fingers and by the time we got the grant we were already doing talents and then CRISPR. So in both cases we kind of exceeded what was going on. So I think synthetic biology is going to have more and more impact on testing hypotheses and moving genomics as often finamins often quoted to really understand something you have to build things with it. And then in C2 sequencing I think is another one more on the back to the analytic side but working together with the synthetic side is as we build more complex synthetic systems like organoids and organs possibly even for transplant for testing hypotheses or testing drugs you'd like to check if they're the real thing and you'd like to understand how the genome, epigenome plays out in every tissue of the body and it's still there's many things that are limited by the fact that I don't know what all the cells are making. We don't have a cell atlas that's a rallying cry that's happening now. I think it has to be a really good cell atlas that ideally in C2 rather than approximating every cell as an isolated sphere not having any neighbors in C2 you get the non-random distribution of proteins and nucleic acids throughout the cell body what other cells surround it but the morphology is important. So I think those are going to be the two big things synthetic, biology testing variance on those significance that we'll be making into the clinic and you can consider gene therapy as a branch of synthetic biology or a sister of it and then in C2 which could help those two along. Well I think the main barrier to PMI success is sharing of data not the quality but I think quality we need to address the quality I think democratization doesn't necessarily result in lower quality. For example the quality of cell phones is much higher now than when only rich people could afford it. Now there's 7 billion cell phones and 7 billion people roughly not quite one-to-one. I think that raw data can be quite poor and the consensus can be quite good so for example PacBio even though it has the worst quality data, raw data it has the best contigs and so when we sequence a genome from de novo we use PacBio because alumina results in 300 contigs and PacBio will put it into one contig for chromosome right away and the consensus is pretty good. Same thing for the nanopores I think one of their advantages is going to be the long reads so long reads gets you high quality just like haplotyping gets you high quality. Anyway I think it's not quite democratized when everybody's cell phone has a sequencer on it then I'll consider democratized because then you'll be reading out your environment as you walk through life and that will be reported out to the cloud. Again sharing data is the key thing for precision medicine I need to know when I walk into this room everything that's in the air everything that's in my food, allergens pathogens non pathogens etc and that will be and I think we're heading there we're sort of in the thousand dollar genome now completely with interpretation and genetic counseling it'll probably be a hundred dollars soon because of companies like BGI and Aluminous moving in that direction they'll only move as quickly as competition forces and move because they are a monopoly and then the antipores will be pushing both of those out of the way with potentially ten dollar or less. Once it gets to a certain low level it becomes democratized and it's free Google Maps is free, a lot of Google services are free to the consumer some of them don't even require you to look at ads I think we're going there very very quickly now even a thousand dollars you can imagine a lot of companies could make money by making it freely available


I'm a single father with two children. An eight year old boy named Terence, and a five year old boy named Leo. You guys probably ask why I'm a single father. It's actually because my wife died after getting murdered by her ex out of jealousy. It's spring break, and I noticed Leo started acting really odd. Sometimes, he would barely eat, he would get random bruises, etc. I felt concerned, so I took him to the doctor, and the doctor had diagnosed him with cancer. Before he passed away, he wanted to see how Spongebob Squarepants was made. Spongebob was his number one favorite TV show. Every morning, he would go to the Nickelodeon channel to see Spongebob, before he goes to school, since he wakes up at 6am in the morning. Knowing how much Leo wanted to see Spongebob behind the scenes, I went to the Make-A-Wish Foundation to make his biggest wish come true. We went to South Korea to visit the rough draft studios to see how Spongebob was animated, and then visited the Nickelodeon Animation Studio to see how they voiced the characters, made the storyboard, etc. There were even shelves of every Spongebob episode in videotapes with the episode names being labeled. I noticed an odd case on the Season 3 episode shelf labeled, Dark and Heart, with some drawings of pieces of trash. I asked them about that, and they said that it was an episode made to spread awareness about the ocean, and how we should be more careful about pollution in the sea, but the writers took the wrong turn, and went too far to the point where it was deemed too dark to be aired and released. I thought that it probably wouldn't be as dark as they said, right? I learned the truth the hard way. In case it was true, I decided to watch it alone to avoid traumatizing anybody wanting to join me. Luckily, I found a room with a TV and a built-in DVD player. Now I can watch the episode, while Leo and Terrence are in the middle of a Spongebob behind the scene store. After I put the DVD in, the Spongebob episode started, and so did my journey of the pollution episode that will soon pollute my innocence. We were greeted with the same old intro from every Spongebob episode. We got the talking pirate in a painting, Spongebob playing the recorder on his nose, and the title card of, You Best It, Dark and Heart. Because everything looked normal, I thought it was just a silly joke to scare me, and that in reality, the episode just suffered the same fate as the Spongebob who could fly, but with no plan to air on TV. Maybe it was exclusive to an aquarium, or part of a perk if you donate to some motion charity. Man, I was wrong. It started off outside, zooming into Spongebob's pineapple, and fading into his bedroom. Inside, Spongebob seems to be shaking in fear, with his eyes closed, like he was having a nightmare. Some videos like the Boston Tea Party, an oil spill, and a sea turtle choking on one a part of a six-pack ring, appear for a couple seconds. He woke up before his alarm went off, and after a couple of seconds when he looks at his clock, the 745th horn wake him up grand alarm clock, or what Spongebob calls, Alarmy, goes off while his face looked like he was looking outside a window at a car on a highway. He felt like he was in a strong windy blizzard climbing up a mountain, using all his grip to avoid getting blown away to turn off his alarm. Soon, Spongebob felt relieved that it was all just a nightmare, and he continued his life like nothing happened. Changing into his clothes and going on a walk to his favorite job, the Krusty Krab. He noticed Mr. Krab's outside, next to lots of floating trash, from water bottles, to pictures of milk. He has buckets of paint and some tubes of glue with him. When Spongebob asked his boss what he was doing, he told him that he wanted to build a large statue of a Krabby Pappy out of trash, and that hiring a sculptor to build a statue out of stone was too expensive. Spongebob walks in and notices customers arguing about random objects inside their Krabby Pappies. One said that a bottle cap was inside his burger. Another said that they had a whole slipper inside.  When Spongebob asks the customer what he's saying, the customer grabs out a piece of paper and a pencil, and wrote to Spongebob, I can't even say anything because there was glue in my Krabby Pappy, and while I was eating, that same thing glued my mouth shut. The customer started chanting the word, refunded, while holding a sign with the same word. The screen goes to Mr. Krabs with his statue finished when his money senses start tingling. The word refund goes from one ear to the other, one letter each. Mr. Krabs dashes inside the restaurant and asks what's wrong. They started yelling at Mr. Krabs, opening their half-eaten Krabby Pappies to reveal random objects inside while yelling out in unison. Explain this. I chuckled a bit, especially on that one scene when a customer reveals an entire real-life scuba diver inside a Krabby Pappy, as the diver swims away, looking embarrassed. Mr. Krabs starts to demand Spongebob to cook the Krabby Pappies, clean and tasty. Spongebob goes into the kitchen to try to make a Krabby Pappy, but it was full of trash. He tells Mr. Krabs about the trash in the kitchen.  The screen transitions to Spongebob and Squidward in the middle of a walk. It looks like the sun is going down, and Squidward complains about how they were walking for hours to find nothing, and that he's going home, throwing the hat on the floor really hard. After Squidward leaves, Spongebob feels upset, wondering how he's gonna solve this mess. Then, he got the idea to visit Sandy, to see if she can find the origin of all the trash. As Spongebob starts running, the screen transitions to the inside Sandy's treat-aum, with Sandy working on something. Spongebob kicks the door open, while panting like he ran a marathon. He tells Sandy about the trash, and that he needs her help to find out where all of the trash is coming from. Sandy tells Spongebob that she was working on an invention, to try to get rid of the trash. During the middle of Sandy talking to Spongebob, the camera pans to the window, as the conversation gets quieter. Some trash bumped into the glass of the treat-aum, and the debris from one of the trash cracked the glass. The camera goes back to Spongebob and Sandy, and they hear a crack. They looked around and noticed the crack on the window. The crack started getting bigger and bigger, before it fully broke and all the water got in. Spongebob quickly takes off his water helmet, pouring all the water out, and gives it to Sandy so she can breathe, while the flood fills up the treat-aum. After the flood, Sandy told Spongebob that she might have to pause on developing something to get rid of the trash, to repair the treat-aum. It's now night time, and Spongebob has found an empty city that looks pretty unerving. He checks the buildings, and they're all empty. Although, he did check a farm, only to find Patrick eating grass, and he told Spongebob that some random dude told him to eat some greens, and he ate grass because grass was green. He was about to leave town, when he noticed something that he hadn't seen before. There was a man with a hat, his identity covered with his silhouette, with a bubble pipe in his mouth. He was sitting in a rocking chair, reading an old newspaper. Next to a rocking chair, shows a gasoline bottle next to him. His eyes can only be seen, which instead of normal eyes, seem to be right-angled triangles. He peeks out of the newspaper, and notices Spongebob. Hello, Spongebob. He said in a deep voice. He sounded like he smoked five packs of cigarettes. Spongebob said, Ah, my name's Spongebob. What's your name? The hat dude then says, Spongebob, Spongebob, whatever. Also, my name's not important. Spongebob starts asking him if he knows where the trash comes from. The hat guy responds, I do know. They're coming. Spongebob's confused. Who's coming? Once you see them, you'll know. The hat guy starts drinking gasoline, until he dies of gasoline intoxication. Then, three fat-like objects, that look like larger versions of common ocean garbage appear. One is an empty water bottle with sharp teeth. Another is a ripped-off food wrapper, with only one eye, a golden tooth, and a spiked bat. The last one is a grocery bag with a scar on an arm, and some broken teeth. The food wrapper's arm reaches out on the inside of the wrap, grabbing a crabby patty, then puts the fist between the bones, the feelings shooting off the burger and getting Spongebob's face. Was that supposed to represent giving Spongebob a knuckle sandwich? The water bottle says, We are the Garanduking. Spongebob looks at the gang, appalled. You were the ones putting random trash in Bikini Bottom? Spongebob asked. The water bottle chuckled. You really are smart for a kitchen sponge. The water bottle starts spitting water on Spongebob, as hard as a pressure washer. Spongebob grabs the ketchup and mustard bottles out of his pockets, and shoots them into the floor for the Garbanzo Gang to trip and fall. The water bottle faintly opens his mouth, like he was breathing his last breath, and then, a dark void starts to come out of his mouth, spreading in every direction it goes, with every object it touches disintegrating into nothingness. Spongebob sprints back into the city he lived in, but it's empty. No people know nothing. Just houses. The void starts catching up, and corners Spongebob. He starts to hallucinate his friends getting harmed by the trash. Spongebob clenches his teeth, and screams in agony. As he disintegrates, with the scream echoing into darkness, the screen fades into Spongebob facing away, crying in the corner while sitting down. Some messages appear saying, 8 x 3 million tons of plastic are dumped into the ocean. Over 1 million marine animals die of plastic debris in the ocean each year. If you don't want the ocean to end up like Spongebob here, stop the pollution. It fades to the credits, but the music was different. Instead of the normal credits music playing, a music box played the same tune of the credits, and the musical scale seemed somewhere between oddly disturbing, too sad. The music aside, the credits play like normal, until we get to the United Plankton Pictures Incorporated logo. Instead of the little happy plankton-like creatures smiling while holding hands, they're all lying down, looking like they're dead. There were some piles of plastic somewhere between them, and in front of all the creatures, there's a small shadow of the garbanzo gang, then it faded to the Nickelodeon logo. I was disturbed, yet surprised. I've seen dark episodes like Are You Happy Now, One Course Meal, whatever, but darkened heart was on a whole new level, and the fact that my youngest son got cancer didn't help. I got Leo into chemotherapy, and I had hope for his survival. Sometimes, I think it's called darkened heart for a reason, and that the name was supposed to point to us. We were the ones who had a darkened heart. We were the ones throwing trash into the ocean. We were the ones putting the aquatic species in danger.


Kia ora Minister Curran, thank you so much for your time and there's a lot of good messages in there for us about collaboration, digitisation and the difference we can make for New Zealanders. The Minister and the Mayor have done such a wonderful job of welcoming you all here and setting the energy and tone for this conference. Andy, if there's a labour of love for a small committee scattered around the country, and I'm still pinching myself that I'm standing here and talking to you all right now, and that we have amazing keynotes, Andrea, Kea, Harkinwall, Claire, Ariana, Pia, who have travelled far and wide and not so far in some cases to teach and learn and share their knowledge and experience and expertise. I want to acknowledge Matariki and Ashley for their support with the mihi whakatau this morning and Ashley's korero for us. It was beautiful, thank you. Now to some housekeeping. Emergency procedures. If you hear continuous alarms, please leave the building quickly and calmly and there'll be people showing us the way out, the assembly point is in front of Te Papa or over in Waitangi Park. If the earth starts to shake, which it won't in touch with, please drop, cover and hold, do the turtle drop and wait, wait instructions, keep away from the windows and wait instructions. There are bathrooms and they're well signposted. There's some just out here and around there but you'll find them all around the building. If you have cell phones, which you all probably do because it's the National Digital Forum, have them on silent if you haven't already, when you're in soundings, that is. If you have food will be served up in Oceania, so up and around, it's well signposted and follow the crowds during morning tea and food shouldn't be taken out of that room because we are in a museum after all. If you use the car park today, you can get your car park, you can get a discount and you can get that your card validated at the registration desk. Wifi, probably should have mentioned that first, right? Te Papa events is the wifi signal and events is the password. We are being filmed. Hi out there, hello to the rest of New Zealand watching NDF and being live streamed as well in this room throughout the conference. So something really important, if you're speaking or if you're asking a question, make sure you've got a mic in front of you because we need the people that are around New Zealand hearing you as well. And it's being recorded too. Sponsors, without our sponsors, you would not be here. I want to acknowledge our car responses to Papa Tongarewa and Manatū Taunga Ministry for Culture and Heritage, to Puna Mataranga o Aotearoa National Library of New Zealand, Tāmaki Painga, Hira Auckland War Memorial Museum, who put some investment into this conference in lots of different ways, not least a lot of the conference committee work for those organisations and there's quite a bit of time that we spend making this happen. So thank you so much. Other sponsors, we've got Internet New Zealand in the house and they have sponsored three people who would not otherwise be here to be in this room. Thank you, Internet New Zealand. They're also sponsoring our drinks tonight, so awesome. Story Box has sponsored the ice creams. Hey, there's ice creams again and thank you Story Box. Go have a chat to them, they're next to the ice cream stand. The DAS Art Museum has supported some of Care's Travel to be here today, Care Winesmith's Travel. And Museums Aotearoa have also sponsored some people who wouldn't otherwise be here to be here and that's so important that we get such a wide, diverse range of people here.  So we've got Gavin and the team at Boost. We've got Andy and the team at New Zealand Micrographic Services, NZMS. We've got Glen and his team from Authentic Limited with MyTourism Curtis, Paul Rowe and Vernon Systems and his team. Marsha from Property 3D with her team and Catherine and the Catalyst team are all part of the exhibit as well as our Kodi supporters have got. So please take the time to go and check out what they've got to show us. Finally, the program is all live and up to date on SHED. So ndf2017.shed.com or go to the ndf website and it's live there. That's the latest program info. There's a couple of changes which I'll cover a little bit later. If you don't have a device, we've got printouts for you at the registration desk and there's some large posters around the building as well. But if you are looking at what's happening next, go to SHED and check it out. Now Pia. I first heard about Pia Andrews. Now it was war, now Andrews, Pia Andrews back in 2011 when the late and much missed Julian Carver told me about her. But I wasn't expecting that to happen. We were looking for judges for the mix and mash competition which was, do you remember that? You tried to encourage people to open up their data and content for reuse and he said, if you want someone who's doing great stuff in this space, there's Pia War in Canberra. She's doing amazing things and she's leading the charge in the open data space and particularly in open government and she's making good shit happen. And here we are six or so years later and she's living in Wellington, working for the Department of Internal Affairs as a service integration lead and she's making really good shit happen for the people of New Zealand and we've got her here today to talk to you about that. So today Pia's going to take us through an exploration of paradigm shifts, considerations for the future and the importance of grasping the opportunity before we accidentally reinvent the past. Pia's appearance today is supported by Tamaki Paingahira, the Auckland War Memorial Museum. Thank you again to our car sponsor and Kia ora Pia, welcome, come on up.