All right, so this is the the last full lecture. It's not super long. It's quite short. So we're just going to, and this is one of the shorter, thankfully, because we only got, when do we end? Five? Okay, so we've got an hour and 20 minutes, just maybe 10 slides in this lecture. And then there's a very brief introduction to the last hands-on exercise. And it's also one of the shorter ones, although this is a pretty big topic, so we're really not doing a really deep dive on it, just sort of getting your feet wet a bit. And I think Brian's going to talk a little bit more about this stuff tomorrow as well, sort of building on some of the things that we're talking about here today. So really this, this module is all about running string tie in a few additional modes that facilitate transcript and isoform discovery. So they're going to, these commands are going to be very familiar to what you already did to do your expression abundance estimation. We're just going to run string tie in slightly different ways to sort of improve its ability to construct previously undiscovered transcripts and alternative isoforms of particular genes. This still does require a reference genome, so the more reference-free stuff is going to come tomorrow. Just to kind of circle back and review the central dogma here, so so far we've been talking about these mRNAs basically being converted to CDNA and then fragmented and sequencing those things by RNA-seq. Alternative splicing is really all about the previous step here where we have single-stranded pre-MRNA molecules that are being spliced together by removing the entrants and stitching together the exons. And of course for many genes and many eukaryotes, there's multiple ways to do this and this manifests as alternative isoforms that differ in the combination of exons they use or the boundaries of some of their exons. There's a number of sort of classic forms of alternative expressions, so starting with just a simple transcript model at the top that's got three exons and two entrants and is being transcribed into an mRNA with the three exons. There's sort of seven modes of alternative expression that are listed on this slide in the next one. The first one is alternative transcript initiation where we've got two possible first exons being used. So in this case you have a transcript with the three exons being used or you might use a different transcript initiation site downstream and wind up with a transcript that just has two exons. And I think we're going to actually find a few examples like that in the data that you guys have been analyzing. And then alternative splicing takes several forms. You can have cassette exon skipping, which is shown here where there's sort of two paths through the set of exons where exon two in this case is either skipped or included. Alternative donor site usage. So this is where the three prime end of an exon gets longer or shorter. So a different slice site of the same exon is used. Similarly we can have alternative three prime slice sites or alternative acceptor site usage where the three prime end of the intron is spliced differently. And it just gives you potentially a very subtle difference in the transcript structure or it could be quite dramatic depending on how much more or less of the exon is included. It's kind of rare but sometimes we'll have mutually exclusive exons where you can have two transcripts that each have the same total number of exons but they're using alternative exons in the middle. Sort of in this case exon 2A or exon 2B and not both. And then the last one that's sort of in the alternative splicing category is just intron retention where the entire intron is retained and effectively it becomes one large exon instead of having exons two and three. You just have a really large exon two. And then finally just like you can have alternative transcript initiation at the beginning of the gene you can have alternative polyadenylation that gives you different exons at the three prime end of the gene. Giving you shorter or longer transcripts. The following site is just kind of a reference material. This is from a review that looked at a whole bunch of different tools and methods for studying splicing by RNA seek. And there's also additional references in the wiki for some of these tools. This is starting to be a few years out of date now but we're trying to keep the tables up to date as new tools are released. Just zooming in a little bit on the transcript reconstruction part of this which is what we're going to focus on in the hands on tutorial. I'm pointing out just a few sort of bio stars post where there's been some sort of ongoing discussion of tools and methods for alternative splicing detection and analysis in various categories here. And just to kind of summarize and put this in context. There's been quite a few approaches over the years that have been used to characterize alternative mRNA form expression. RNA seek has really become the most popular way. But there is still probably application for some of the other techniques. So someone mentioned pack bio sequencing here if you really want to understand the full length structure of a transcript you might decide to take advantage of the longer reads that you get out of the pack bio. And maybe in a targeted sense try to sequence some full length RNA sequences on the pack bio or much longer. People are still doing CDNA synthesis and sequencing where you really need that gold standard or where you want to actually produce the reagent where you actually have a physical copy of the transcript in a plasmid that you can use for further experimentation. There's just a ton of existing data out there that you may also want to leverage. So just to review this is we're sort of going to take a little bit of a step back and we're going to go back to the steps that we're running at the beginning of the expression module and basically rerun string tie with some different options. And then we'll set up the files that you would need to do ballgown analysis again as well. And then we're going to look at some examples in IGV so you can kind of see what this looks like in our example data. So just to briefly introduce the hands on component of this module. So we've already been running string tie in what we've called reference only mode. Now we're going to try running it reference guided and de novo modes. And then we're going to learn how to use cuff merge to combine transcript domes from multiple actually string tie runs that should say and compare the assembled transcripts to known transcripts. And then we're going to perform differential splicing analysis or at least do the setup that you would need to do for differential splicing analysis with ballgown. It's really exactly the way you already did it. You're just going to have GTF files that were generated in a slightly different way and expression estimates that were generated in a slightly different way. We're going to examine some junction counts with a sort of helper tool that we developed called reg tools. And then we're going to look at some junction splicing files or outputs in IGV along with the string tie assembled transcripts that we were able to generate. Okay, so how do we run string tie in rough guided and de novo modes? Basically we're going to do this by taking advantage of some additional options. And it all has to do with basically how and whether we take advantage of the GTF file of known transcript annotations that we've been using. So we're using this GTF file in several places, so it kind of has been we're sort of keep coming back to it over and over again. Remember during the actual indexing of our genome before we even did our alignments with high set, we use the GTF to provide the aligner with additional information about exon exon junctions and about the boundaries of exons to help it actually align the RNA secrets against the reference genome. And then we used the GTF file and string tie by supplying with the dash G option to where we're going to do this to run string tie in the so-called reference guided mode. So this is a mode where previously in the reference only mode we basically were telling string tie give me expression estimates for all of these transcripts. Now we're going to run string tie and we're going to say, yes, do that, but also look for novel transcripts, but use the GTF file as a guide. So use whatever information is in there to help you figure out where transcripts ought to be. But if you happen to see any novel exons or novel exon boundaries, then we want to know about that. And then the reference, the de novo mode, we're basically not going to give it a GTF at all. So it doesn't even know about the transcript structures at all. It's just going to look at the RNA seek data and at the reference genome and it's going to try to figure out what the structure of transcripts ought to be. So this is a much more challenging problem now. It has no prior knowledge of the boundaries of the exons or how the exons are connected in this set of known transcripts, which for human is a really rich source of information. So we have these really great, you know, pretty highly vetted annotations for the human transcript dome that have been built over the years with really high quality data from ESD sequencing and CDNA sequencing and so on. It's going to try to do everything that it's been doing already without the advantage of having any of that information, which is a big difference from the reference only mode where we use this E option to say, basically give me estimates for these known transcripts. And then after alignment, we're going to create a junctions bed file. So this is shown, an example is shown at the bottom here. This is just a way of summarizing what exon-exon connections were found in the RNA seek data. So all of the reads that happened to span across an exon boundary, we're just going to pull out all of those cases and summarize them specifically and sort of ignore the RNA seek reads that fall within exons. Just to get a sort of a map of the connectivity of all of the exons. And when we look at this data in IGV, we'll get a view that looks something like this where for each junction that was observed, we'll display a little arc that shows basically where the read goes from and to spanning across an intron. And then the number of reads that supported each unique combination of two exons. So each of these arcs corresponds to a combination of one exon to another exon. And the sort of thickness of the arc is a representation of how many reads there were that supported that particular exon-exon combination. And this is an example where you can actually see evidence for what appears to be a novel exon. So you can see that most of these arcs correspond to known exons. So we have a gene model up here that would have been represented in our GTF file for a particular gene. And you've got pairs of exons. And then you've got these arcs that seem to correspond to the edges of all of these known exons as we go from the beginning of the gene to the end of the gene. They seem to really match up really nicely. But then when we get over to this intron, we can see, yes, there is a junction that spans from this expected exon all the way over to this expected exon. But we're also seeing two additional exon-exon junctions that are sort of going into the middle of the intron and landing actually kind of close to each other. And this can be really suggestive that there's actually a novel exon somewhere around here where its acceptor site is here and its donor site is there. So we could zoom in on the RNA-seq data in that region and we could try to see how much evidence is there that there's actually potentially a novel exon there. Yeah. Can we separate the graph with three rows? Are they all the same difference? Yeah, they're just being spread out for display purposes. So in IGV, you can compact them so they're all on one line. But then sometimes it gets a little bit hard to see them properly when you have instances like this where there's exon skipping and one of them is going over top of others. So there's usually this option to kind of expand it all out so that everything is really easy to see. Any other questions on any of that? Okay. We're going to play around a little bit with this tool called cuff merge. So this is really a way to combine transcripts that were predicted from multiple RNA-seq data sets into sort of one unified representation of the transcriptome.  So maybe you have five conditions that you're interested in comparing for biological reasons. So you're processing them separately because you want to ultimately compare those five. So then when it comes to predicting the structure of transcripts, you might decide that, you know what, let's just pool all that data together because for the purpose of just understanding what transcripts are there, we just want more data available. And so cuff merge is a way of basically say sort of amalgamating the transcript prediction information from those five runs into a single run. And then you could use that as a new reference transcriptome and go back and redo transcript estimation from each of your five individual conditions using knowledge about the structure of the transcripts that was gleaned by looking at all of the data together. We're going to do some comparisons on our own in IGV of the merged ETFs that come from each of these different string time modes. This is just showing a couple examples. I'm just going to skip over that because we're going to look at some examples ourselves using our own data and that will be more fun than looking at it in slide form.


Our Tory government's approach to COVID has been consistently undermined by ignorance, denial and appeasement of the right-wing libertarians who occupy the party's back benches. Each one of these deficiencies were on show in this ridiculous answer from Sajid Javed on Sky News. How many of you were wearing masks? You can take a look at the picture that we've got. The number 10 released yesterday, they chose this picture. There's basically, well, 38 to 40 people in that room, poorly ventilated. Not one person is wearing a mask on the day that you advise us to wear masks in situations like that. And that is perfectly consistent with what the Prime Minister said yesterday and what I said yesterday, because what we said was that people should consider wearing masks in crowded places when they are with strangers, when they are with people that they are not normally spending time with. So it's very conservative back benches to be wearing masks at PMQs later today. No, they're not strangers. The sort of back benches, whether they're in parliament, in the chamber itself or in the other meeting rooms and things. You don't catch COVID for your friends. You have to take measures that are appropriate for the prevalence of COVID at the time. That guy is the health secretary. He's the health secretary. He's the most important person in the country when it comes to public health. And he's going on national television and saying, you don't have to worry about catching COVID if you're among friends. And now I can imagine you saying, oh, you've got a gathering, you've got a party, it's not really going to work if you're all wearing masks. He's talking about the House of Commons. That's 650 people. The House of Commons would function perfectly if everyone other than the speaker and the people who were speaking were wearing masks. There's no reason not to wear it other than ideology. There are a number of ways in which I think that clip is characteristic of everything that's been wrong with the toy response to COVID over the past 18 months.  As I say, you can catch COVID from strangers. COVID doesn't care. If Tory MPs went to the same debating society, if they trashed the same restaurants at university and public school, it doesn't care. They can still pass COVID to each other. Secondly, what this does is show a government completely in denial. Javid there said, we have to take measures that are appropriate to the prevalence of COVID in society. That implies we currently have low rates. But let's see how we compare to our European neighbours. These are new confirmed COVID-19 cases per million people in Britain and in our European neighbours. We are running at about 500 daily cases per million people. France, the Netherlands, Germany, Italy and Spain are all under 150. Now, when faced with these kind of comparisons, what you'll often hear is, we've got higher vaccination rates, which means that a case is less serious than it is there. Actually, that used to be the case. Now, most of those countries have caught up or overtaken us when it comes to the number of people vaccinated. It's also worth noting all those countries have mask mandates. This is another situation of complete denial and exceptionalism from the government, which is putting people at risk for no good reason. If we can limit transmission in a relatively socially cost-free way, just do it. They're refusing to do it. The final thing this is why it represents everything that is wrong with our government's strategy when it comes to policy making, is that that was our health secretary. Now, he should be making policies that protect our health, communicating the latest up-to-date science on COVID-19. Yet, instead, he's essentially lying to the public on an issue of public health, all to justify the pigheadedness of his extremist backbenches. He sees the backbenches, they refuse to wear masks. Instead of him going on television and saying, how do I communicate the best up-to-date science, he's saying, how do I go on television and justify the idiotic behaviour of the backbenches in my party. Now, if you want an idea of how strongly these adult babies on the Tory backbenches feel about masks, just look at this moment from Tuesday's announcement about our so-called Plan B. We have prepared a Plan B of a contingency measures that we can call upon only if they are needed and supported by the data to prevent unsustainable pressure on the NHS. These measures would be communicating clearly and urgently to the public the need for caution, legally mandating face coverings in certain settings. Now, like Sajid Javid there had literally just said, if we were in a situation where the NHS is under severe strain, I mean, it is already under severe strain, but he's, I'm presuming he's imagining it's about to collapse, a Tory backbencher MP has heard that in that extreme scenario people are going to be asked to wear masks and he, you know, he's at work that now, like it's, it's so pathetic, but it's also incredibly influential because it seems to be dictating government policy. Dalia, I want to bring you, you in on this. How significant do you think these extremist backbenches are? Am I in a way letting Boris Johnson and Sajid Javid off the hook because they are also just as extremist as that person sort of screaming now in the background? Would it have been Boris Johnson sort of go, no, if it was Theresa May who was still Prime Minister announcing a sensible policy such as this? It's very, very hard to say because I think a lot of people that are, have been close to Boris Johnson have often said that the only thing he cares about is, you know, the, the Murdoch papers and, and the Tory backbenches. But it makes you think, what are you scared that the Tory backbenchers are going to do? Like what kind of threat do these, you know, I mean, that kind of dinosaur style croaked the idea of wearing a mask doesn't suggest to me that this is some big, early threatening person that I should be really scared of. It sounds like someone who's just sort of a bit of a coward snowflake, really. So I don't really know what they're worried, you know, that if the Tory backbenches are sort of, you know, coming to you, what exactly they're going to do. But I think that, you know, obviously that, that interview where Sadja Javid said that thing about, you know, not, you can't catch it from people that you know is absolutely, it's actually the precise and exact opposite. I feel like you're actually more likely to catch it from someone that you know, because you're more likely to, you know, being close contact with them, you're more likely to be an ongoing contact with them, you're more likely to be, you know, hugging them and being sort of near them without a mask on. I mean, I don't, don't quote me on that. I don't know the exact statistics, but it strikes me as probably quite likely. I know that when the, in the, the sort of the last massive uptick in infections over winter during that Christmas period, a big reason for that was secondary schools.  Like COVID isn't something that you give to someone because you don't like them.  And I also, to finish this off, and this is something that I don't think we're picking up on enough. The fact that one of the plan B strategies, which, you know, is going to kick in if, you know, the NHS is on the brink of collapse. And, you know, however many deaths per day is to use caution. I mean, this government has not learned a single thing. What, what does that mean? Throw that in the bin. I'm fed up of it. Like that's just, what does it mean to exercise caution when you haven't given people concrete steps on how to be cautious? It's just absolutely absurd. And it just shows to me again. And this is where I think a bit more of the blame should go on to that front, the front bench and the decision makers is because I think it's not, it's rooted partly in political expediency, but I think it's rooted more in actually being quite ideologically consistent with the idea that, you know, mandating people to wear a mask is like some kind of violation of individual libertarianism, which is of course absurd. I think that's such a good point. I hadn't picked up on that actually, that plan B, if the hospitals are about to collapse, will urge the public to use caution. Like, why don't you urge us to use caution now? You know, that doesn't, that seems like that should, that shouldn't even be plan A, that should just be like a, a given. Just a plan. Until the pandemic is over, like the starting point should probably be like, use some caution. You know, very absurd. It's absurd. And it is, it's deeply ideological. And it has cost tens of thousands of lives. Right. We would have had a much, much lower death toll if instead of following his back benches and following his own, I don't know, was like right-wing libertarian instincts, I suppose. Boris Johnson had just literally followed science, right? We wouldn't have been, what the Julie Hartley Brewers of the world will say, if we follow the science, it would have been endless lockdowns. Not at all. That's not really what the scientists were ever suggesting, what they were mainly suggesting, because they do actually seem to care about freedoms, despite this whole like, oh, they're merchants of doom who just want to increase their control. That's the Dan Woodton line on GB news. They, they do actually tend to seem quite interested in, in balancing freedoms with safety, which means that they are highlighting things such as mass, such as certification, these things which won't significantly impact our lives, but which will keep the virus under control and ultimately avoid lockdowns. That's what Patrick balance kept saying. If we use these simple options now, then we won't have lockdowns in future. So it's not Boris Johnson on the side of freedom versus the scientists on the side of totalitarianism. It's Boris Johnson on the side of, I suppose, this sort of anti science irrationalism, which over and over again puts off the inevitable as a tool of party management. So he's like, oh, they won't be able to oppose mass mandates if the hospitals are about to collapse. So I'll wait until the hospitals are collapsing. It's the worst way to do public health imaginable.


There's something that I see a lot of you doing Which is you're waiting until you are ready before you do that launch before you make that video before you publish that book or announce the course or Whatever it is that you want to get out into the world so that you can Create an authentic business Visibility for your business, etc. You are waiting too long before you are ready and One of my dear clients mentioned the the thin line between readiness and procrastination And how do you know if you are not yet ready or whether you are procrastinating? Let's talk about this First of all you probably grew up in a world where businesses were very expensive to launch You had to manufacture in your most businesses right before the digital world had to manufacture a product Which means a lot of cost raw materials, you know a manufacturing line, you know getting it on retail shelves There's limited retail shelf space, so it was expend. It used to be risky and expensive to launch anything That's why Businesses had to do a lot of preparation had to be super clear about what they were doing had to minimize all the risks before they launched Well, you're not manufacturing a product and If you do these days someone else like Amazon will manufacture it for you And you just have to give them the specs right now many well Amazon manufacturers books all the time, right? You just have to give them upload your draft and they do the print on demand and you don't put any money out If somebody buys then they print it and they give you the profit so so everything in the digital world The lot the risk of launching a business is way lower Okay, and also Even the risk of a first impression being bad is way lower Because with the digital marketing Even if they didn't like what they first saw from you They will see you again and again and again and they will first of all they probably Forget so these days a bad first impression is not like I am never going to watch George's video again I am never going to read his article again. It's more like that wasn't that interesting to me I'll just and then you'll see my another video for me Not that interesting a third video you see for me Oh, that was kind of interesting and then you watch it or you watch part of it And then you have a better impression me so in other words nowadays It's not like oh, I'm so scared that if I don't make the first good first impression That the person is going to leave forever that is not true anymore It's basically the bad first impression is like meh Okay, and then you know and then they're they're holding nothing against you Unless you're purposely trying to offend people Right, which you're you're not and if you accidentally offend somebody by the way, they're not the right ideal client for you Anyway, they're not the ideal reader your ideal reader is not going to be offended by what you do authentically That's the definition of an ideal reader or ideal viewer is they are Naturally more resonant they are naturally more likely to accept and enjoy How you are authentically than somebody else? okay, so Let's only make things for our ideal audience of whom there are probably millions out there with Facebook for example with Facebook ads you can reach 250 million people in the United States alone if You're in Europe Canada or if you want to reach Europeans or Canadians or Australians or Singaporeans or you know name name whatever country you want South Africans, right? You now expand your reach by tens of millions of people hundreds of millions of people actually sorry so Do not be afraid to make a bad First impression because nowadays a bad first impression is only meh. I'll just wait until I find something interesting That's all that's it. So what are you so afraid of so in other words? the readiness is Really, I'm almost certain that if you're seeing saying I'm not ready. It's because you're procrastinating I am almost certain of it. It's like 90% chance and if you disagree with me Please comment below and let me know why you're not ready. Okay, so let me let me tell you why I Feel so certain about this To I never feel ready Okay, I didn't feel ready before I pressed the record button because I said my god I could have I can really be brainstorming around this topic for another few hours before I get I feel ready to Turn on the camera and start talking. I don't feel ready I didn't feel ready my my first ever video and I still don't feel ready now I'm probably I think I'm getting close video 1,000. I still don't feel ready and I'm gonna write an article later about this. I don't feel ready to write it. I'm scared It's gonna be a blank screen. I have a few notes scribbled down and how do I make it sound amazing for you? Maybe you won't like it. Maybe you don't think it's elegant enough. I don't I don't know But here's the thing. I've noticed something Once I do it when I'm in the doing of it. I feel more ready like right now Obviously the camera is already going. I've already pressed record. It's already live I feel more ready than I did before that's the reality of it You don't feel ready when you start and you just have to force yourself to start now the forcing of yourself Doesn't have to be pain and suffering and like hustle and grinding teeth and pulling self up by bootstraps those kinds of metaphors Forcing yourself to start for me is like this It's kind of like I turned fear into excitement. That's what I want you to practice Turn your fear, which is what your lack of readiness is is your fear. That's it And that's really the core lesson of creative self-expression. That's the core lesson of creating an authentic business Turning fear into love Turning fear into passion Turning fear into service turning fear into curiosity turning fear into self-exploration turning fear into Abandoned sort of like a a reckless abandon of the child And it's really not just an authentic business lesson. It's also a lesson for a truly self-actualized life It's turning fear into love And when you say I'm not ready you are giving in to fear. That's it You're giving into fear.  Will you turn fear into love now will you turn fear into a child like you know? Jesus says you know You won't enter the kingdom of heaven unless you're like this little child now Of course people have interpreted that a million ways over the years, but let me do Some blasphemy now and interpret it in this way You won't enter the kingdom of heaven unless you have the reckless abandonment and curiosity and love of a child You know and the innocence and the humility Right That's also right when you say when you say you're not ready You're not being humble Right, you're saying ironically you think I'm being humble. I'm not ready. No, no, you're not being humble You are protecting your ego right when true humility is Offering something that may or may not be ready and asking the market. What do you think? Do you think this is ready? So here's the key You're not the one to tell you if you're ready or not the market tells you if you are ready so a famous quote by the founder of linked in who has of course not just linked in he has also Co-founded several other successful companies been a very successful investor. He said this If you are not embarrassed by the first version of your product you have launched too late If you're not embarrassed by the first version of your product you have launched too late I'm kind of embarrassed by this video, but that means I haven't launched too late because You know, I'm learning as I go, right? I'm embarrassed slightly by every article that I put out there I'm embarrassed slightly Sometimes moderately by every book that I put out there's honestly honestly. I'm embarrassed by every course I teach And and here's the here's the here's the reality of it Once you look back, you will always be embarrassed That's really what's and I want to challenge you to be slightly embarrassed when you start I'm not saying to be like mortified and like this is the worst thing I've ever done I'm not saying that and that's not what LinkedIn founders saying to he actually wrote a wonderful article to clarify what he meant And I will and I will link that article in the notes of this video too But he said listen the reality is that when you look back you will always be embarrassed by what you just did because you Because you you'll be learning in the doing of it and so you look back so I could have done that better And so that embarrassment is normal Did you think that that embarrassment meant that oh, I wasn't ready and I shouldn't do it sooner next time? No We learn by doing we truly learn By experimenting not by waiting until we're ready. So here's what's going to happen if you wait till you're ready You will always do things too late And you will always run out of time Okay, I I've known people who didn't start until they were in their 70s They wish they started in their 50s Or their 60s even they wish they started in their 60s But they didn't start until their 70s and they're like god I really shouldn't have waited until I was ready because you'll you'll never feel ready. You'll never feel ready Honestly, and when you wait until you're ready, it means that you've you're too late because you have you've run out of money Okay, the people who are like Oh, I've got a year now to start my business right because maybe I have a savings of a year now Just to run my business and they wait until Month 10 to finally launch their thing and now they only have two months left to learn or to try to Build an audience before they run out of money and have to go get a job again No If you have a year Say it to to start a business you should be launching in year in month minus one if you have if you're gonna say, okay January Out of a job to start a business You should be starting now in october of 2018 while you're still in a job You should be building an audience Not wait till january It takes longer than a year to start a business Because it takes longer than a year typically to build an audience Who care enough about your about you and about your energy and about your thoughts and about your Care enough about your you know your presence to even give your product maybe a Right and also it takes about a year now I'm not going to say it takes a year some of you might take only three months because you work Dilligently you are experimental in mindset, etc It might only take you three months to discover that product that takes off that service that takes off Okay, so it doesn't have to take a year But a year is a pretty reasonable bet honestly if I were betting if I were investing in your company and your business and your solo premier business I'd say I'm not going to bet that you're going to succeed in three months a year More more likely I won't probably won't see a return on and as an investor I probably won't see a return until you're two three or four or five But I could see you sustaining your your family in a year, you know, I could see you do that And essentially I feel like I'm investing in my clients I'm not literally investing my money, but I'm investing my my heart and my time and sort of the risk of being their coach so Okay It takes So, okay, so let's let's talk about readiness because there is something about market readiness and something about audience readiness And by the way in this case you're wondering I'm in a different environment today because I'm visiting my mom for a few days and this is her bedroom This is like the only quiet place I could kind of set up a place to to do a video With adequate lighting So here I am in my mom's but so There is market readiness. There is audience readiness Okay, that is that's a real thing. Okay, so I think of business launches in stages. You don't just say George, I've got a service I want to launch and it's and it better be successful the first time I launch it If you tell me that I I realize oh you haven't don't have any business experience. I see that's why you think that Once you get some business experience or if you listen to somebody who does and who has coached By this point hundreds personally one-on-one hundreds of clients actually I think they're in close to a thousand now Um, maybe more I should actually count.  Really that's amazing or you launch your course or you launch your book or you launch your product You pick you got somebody bought it. You know how hard it is to get people to buy anything That's amazing Business experience has shown me and has shown pretty much everybody which is why People like you know successful investors and business founders like read hopman says you got to be embarrassed by the first version of your product Because the real learning happens after you launch The real learning happens after you're promoting your book after you've Published your book after you've launched the course. That's the real learning happens After you've made the video after you've published published that blog post after you've started your facebook business page After you've started your youtube channel.  That's where the stages are. So let me tell you this I talked about market readiness and audience readiness That is that is a better way of talking about readiness. Not do I feel ready? Am I feeling inspired right now? Am I oh god? Am I am I skilled enough? No, let me go take another course Then i'll be good enough That is I've noticed that that's particularly true for a lot of women No, and i'm not being Chauvinistic the studies research has shown that women are more likely to feel insecure By not having enough degrees and certifications and training programs and classes Women you need to know that that is statistically significant That you are more likely than men and so yes all the quotes about you know move fast and break things are from men Usually you know white men right so genetically speaking Maybe white men are more likely to be more entrepreneurial and more experimental mindset And women I don't know if it's particular race or whatever but women are more likely to feel like let me have to I have to get ready And the way I get ready is to read another book take another course Get another certification get another degree then I will be ready So that's what educators and schools They will market to you For with that They will market to your uncertainty and your doubt and your fear that you are not yet ready You got to get this degree first then you'll be ready. Okay. I was An admissions director. I was I marketed a graduate program for just a couple of years So I know what I'm talking about And now I market courses my own courses now. I hope I don't I'm not marketing to your insecurity. I hope I know Please call me out on it if you see many of my marketing copy That's marketing to your insecurity because I do want to change and I still And learning how to change myself towards more authenticity. I think I've come some way But I know I have more to go. So please call me out. I'm serious about that Okay, so There is market readiness. There is audience rated. It's not about your readiness It's about your market and your audience's readiness. So how do we know that we build business and stages We build it and that's why I have a 10-year plan Please go to george cow.com slash blog slash 10-year plan I've written a long blog post detailing my own 10-year plan Uh, I'm building a brand new business. I'm following my own 10-year plan there And this business that I built basically has followed that 10-year plan and I'm in year five basically right now of my 10-year plan I'm already Very good full-time income from this business has have been so for several years But there's more to come and so here's the here the stages Stage one build an audience Build an audience now. I'm assuming that you're wanting to build an authentic business Which I define as a business where your creative self-expression makes you money How would you like that? How would you like to be able to be yourself? In writing or on camera or on a podcast or being yourself in your creative self-expression and people pay you to do it That's an authentic business. That's the 10-year plan Now you make money after as soon as year two you make money and maybe even full-time money in year two We'll see but um depends on how quickly you apply my plan so Step one build an audience. Okay Step two you make your audience an offer and see What their reactions are And if the reactions are nobody bought it, which is normal that nobody buys your first launch Maybe a few people if you're really lucky a few people buy it if you're super lucky You get a lot of people buying it, but it's rare. Like I said after you know Talk with thousands of people by this point entrepreneurs Step one build an audience step two make an offer step three Take the feedback or lack of feedback from your offer if it's lack of feedback You need to do some fan interviews You need to have one-on-one conversations with your audience to say hey, I'm wondering what are you looking for?  That's I call those fan interviews or audience research conversations. That's step three, right? Do audience research conversations after you've launched something that didn't work, which is normal you launched something Why it's a george? Why would you launch something that doesn't work because you don't know if it's gonna work or not? You don't know but you need to expect that it won't work Okay, you need to expect that the first launch is just getting their reactions Planting a seed you're planting seeds as the first maybe the second launch The third launch you've gotten enough reactions and learning from conversations that the third launch probably will work Okay, really because you've been tweaking for a couple times now Okay, and then step four is to scale the thing that works And then you go back to step one whatever the next product you want to launch Build an audience now by this point you already have an audience, but at least you are creating content that is Educating the audience giving context for the next launch for the next product, right? Not like you're pre-launching the whole time, but at least you're giving content That's related to the to the next product, right? Step one step two you launch it see if they like it They probably nobody will probably buy it, but they'll actually give you some comments maybe Or you can talk to them and find out what they're buying instead or what they didn't like about this one And then tweak it step three, you know launch it again It works maybe the second or third time step four you scale it with ads so that you can Eventually have a passive income if you're launching things like courses, okay? For example do it yourself courses and you can have a passive income But you see what I mean like and and so Let's talk about a market and audience readiness The the audience readiness happens as you build an audience and and create content and you notice the reactions to your content You notice. Oh my gosh every time I talk about this. They really like it Oh, so that might be what my first product it should be Should be my first course my first book my first service offering my first coaching package my first You know group program Oh every time I write about that people really like it Oh, maybe that should be my first my next first or next thing I launch So audience readiness is a real thing, but you notice it after you create content and share it in the market Hopefully you're you're using some advertising dollars to promote your content to new people who don't know you yet Because otherwise building an audience is really being slow just with friends and family um So do you see what I mean? I know I've been talking kind of fast, but I Hope that this video will help you get out of the illusion of being ready because Every time you see see yourself saying oh, I don't feel ready I'll remember george said there is no such thing as personal readiness There is only market and audience readiness which happens once I start putting content out there And then I observe their reactions to my content to notice what they're ready for What is my audience ready for and therefore that's what I'm going to promote next Knowing that the first promotion probably won't get any sales But at least it'll plant a seed for them for the next one. I'm going to promote or at least I'll get Oh, that's only what I know if they're going to buy is to put stuff out there Okay, so uh, I hope this is helpful and I hope you will Realize that the risk today And I'll just reiterate what I said in the very beginning You're not You're not flying a plane you're not You know, you don't want to pilot to your airline pilot you get on the flight and the pilot says Well, I'm hope I'm ready to fly this plane Okay, it's not flying a plane plus the pilot also went through stages of readiness The pilot didn't just jump on a plane. So I read I read a book about flying a plane I'm going to go and try to fly this plane now with 100 people on board. No The pilot, you know said am I ready to go to flight school? Oh, no, I'm I'm being dangerous I'm saying, you know, you should go get get another degree. That's what I not what I mean But the pilot went through multiple simulations It's okay. Now I'm ready for the next simulation now I'm ready for the next simulation now. I'm ready for the next simulation Well, now I've done fine with five 10 50 simulations now. I'm obviously ready to fly a plane But that's the same thing with your business each simulation is a launch Each simulation like a pilot is a launch in your business So you need to build your audience in building your audience is like going to flight school Don't get another degree. Please. Please. Please. You don't need I mean if you're building an authentic business about your self-expression creative self-expression You don't need another certification if it's about your creative self-expression Now it's about some skill set that is recognized in the marketplace that people your prospective clients want you to have Well, yes, of course to get a certification in that But you're not doing brain surgery And you're not flying a plane Okay, you are a coach.    You are an author You are somebody who wants to just talk for a living or write for a living Or help people for a living doing something you love to do And it's not, you know In the marketplace for you to have a degree. Please stop Don't get another degree. Don't get a notification.  Do it for fun Do it for yourself enrichment, but not for oh, so then I'm ready to launch my thing Okay, so back to what I said in the beginning Launching a digital business Business now nowadays authentic business. There is no risk. There is no risk of a bad first impression Watch the beginning of this video There is no risk anymore. It's just like meh. I didn't like this video. Ah, I didn't like her product Oh, I didn't like her blog post It's not like oh, I'm never going to read her thing again. I'm never going to watch her again No, no, no They'll just wait for your next thing to see if they like it And then it's sort of like there's you can't go below zero They just will be like meh meh meh. They don't like your thing. They don't like 10 of your posts They don't like 20 videos meh meh meh you're at zero and then you make something they like Oh, now you're at a one now you're at a two Okay, and then now you're at a two as baseline. Ah, they didn't know the next thing you do They don't like they don't like they don't like it.  They'll never dislike you.   So anyway, I hope this is helpful Thanks for those who were able to join me here for this and thanks Linda T. Okai, Sandy, Prem, Rob, Lori, Joseph, Lisa, Cheryl And captain and Pamela, Tina, Annie and Robin, so thank you all. I'm just going to mention a couple comments here. Um So, uh, let's see here Yeah, Lori says things are sharing that even with all that experience you feel scared I feel scared before every video that I make and probably In about the first 25% of the video, I still feel scared By this point, I'm not scared anymore. Now I'm kind of scared of what your reactions are but But still he's even though I feel scared. It's the muscle I've built of ignoring the fear rather than having to somehow You know, figure a solution for the fear. I just kind of ignore it. I try to turn the energy into love and passion and experimentation Yeah, Lori says Yes, fear and excitement are two sides of the same coin often when we think we're scared. We're also excited. It's the same physiological energy that we actually have so we can mentally turn the physiology of Into oh, I'm excited then that means mean I'm excited that must mean there's some passion there Because there is you won't be scared of anything if you didn't care about it right, so Yeah Lori says bottom line if you haven't started yet start now on whatever it is your you feel you're not ready for Yeah, and you know what I remember the stages Whatever You're doing take it to the next stage. Okay, so what I don't mean is oh George says I better launch a business now Even though I'm not ready. I'm gonna I'm gonna borrow a loan of $10,000 from the bank to run facebook ads That's called foolishness. Okay, but what George is saying is take the next stage the next step of something you're not ready for Oh, I'm not ready for video What's the risk? Oh, the risk is that people might hate me.  Okay The ones who are your ideal audience are not going to hate you They're only going to be neutral And the ones who hate you or don't dislike you are not your ideal audience anyway never will be okay, so The next thing you're you're scared about video Writing a blog post building an audience That's probably the next thing and after that launching the first thing and a very simple and lightweight not spending thousands of dollars on That's something a lightweight onward Yeah Prem says it's comforting to know that the perceived need for more training is Statistically significant for women. This is why More and more even in higher education There's more and more women of you know females in higher ed more more and more so than than men That's part of this but also just in adult training adult education. It's it's mostly women More and more so anyway Janko. Thanks for being here as well. I'm gonna let you go. Thanks for Being part of this video and any questions. Let me know below any comments Anything you want me to create a video on that you you would find helpful. Let me know Be well. Have a great day


I know these people's stories. I know about Pusha T saying, yeah, I grew up in a two-parent household as well. I know about Jada Kids being like, yeah, I grew up in a two-parent household as well. Just cause you grew up with that structure don't mean, what's Pusha T favorite thing to rev about selling cocaine?  But for you to have that, and then they still together now, right? Still together, they been married for over 40 years. Awesome. That's big, man. That's huge when you come from that. Just explain to me, coming up as a kid in New Orleans, how was it for you? How was it? So I didn't realize that we lived in a war zone because inside the walls of that house we had, we had a little small town house, you know what I'm saying? Like not anything big or lavish at all, but it was so filled with love and structure that I didn't realize that outside of this house, man, it's a trap house across the street, one house to the right, it's, you know, police tape cause people getting murked and people getting shot all in the neighborhood. Like, this is considered a place where, you know, people with money definitely don't live around here. I never realized any of that until I moved away and went to college because when I moved away, I was, you know, in a different environment and now when I would come back home, I was like, ooh, it feels different when I come back home than what I'm used to up in college, you know what I'm saying? My daddy, blue collar worker, welder, you know what I'm saying? My mom's social worker. So they, I just saw him wake up every day, make an honest living, go to work. My pops worked an hour away from our crib. So he had to wake up at four in the morning and drive an hour each day just to get to work welding in a hundred degree heat, New Orleans heat, you know what I'm saying, all day, every day. And by the time he'd get off of work, like 10 hours later, he coming back, he making it in time for my baseball practice, for my basketball games, you know, still doing this stuff. My pops used to be the dude who wanted to tape everything, video everything. So this wasn't in the era of smartphones and all that.  And he that he's sitting in the stands, players. Yeah, he's sitting, he's sitting in the stands and he, he capturing all the memories for me. So now when I look back at all my high school games, I'm like, dang, I got this on tapes because my daddy, after working 10, 12 hours, you know, dirty, dicky outfit on from welding all day, making it in time because he prioritized that. Like I just seen that's what was normal to me. So that's how it was coming up. It was like a combination of that. And also my life turned out different than a lot of my peers because the school system in New Orleans is not the best. It's set up to where you supposed to go to your district school. Of course. And if you don't have a lot of money, if you live in a poverty stricken area, your district school is trash a lot of times. The district schools that I was supposed to go to, they were, they were not, they were not conducive for raising young black men and women in a, in a loving place that also was academically rigorous. You know what I'm saying? It was just, it was dysfunction and it was like, get it how you live. You know, we're going to have a couple of shining stars that's going to emerge from here, but for the most part, the bar is set really low. That's, that was all my schools, right? But I never went to those schools because my daddy, when I was in kindergarten, they had a school all the way across town by Tulane University and Loyola University. It was called Audubon Montessori. They realized that like that right there, that type of education is free because it's still a public school, but it's also going to show David. That's me, you know, my name is David. It's going to show David a different side of life than what we can provide for him over here, where we live at in the East. They had to drive you to school every morning? The first of all, he had to camp out in a tent overnight to have a chance to enter a lottery to get me into the school because so many people want to go to the school. Right. So there was like the first 100 people have a chance to get into this school. My daddy camped out the night before in a tent on the side walk and he was number six in line. They had a line down the block. You heard me? Like a line to get in heaven or something. It was like, yo, people trying to get here. My daddy got there so early, got me a spot. I hit the lottery, you know what I mean? Hit the lottery, got in that school. That school, that changed my life because I went there from kindergarten to eighth grade. And every day, although I'm living in one type of environment, I'm going to school with black students, white students, Asian students, Hispanic students. I got to walk past million dollar houses. The type of houses that Drew Brees lives in the type of houses like Anthony Davis will be living in when he was playing for the Pelicans. And I got to pass all this up every day. It's on the same block as my school, you know what I mean? This became normal to me to where it's like, oh, I'm not intimidated by white people. I'm not intimidated by other ethnicities thinking they smarter than me. Y'all have more money than me back then. But I was in gifted classes, you know what I mean? So I'm seeing as a young kid like that analyze certain things. Yeah, like the whole the whole concept of idolizing material possessions, like that was that was dead to me. By the time I was in middle school, I was like, man. Yeah, your parents got money so y'all live in this big fancy neighborhood. But when we get in the books, I'm doing better than you in school. You asking me for help. You trying to copy off my paper, you know what I mean? Like it's life experiences that show me like, I'm lacking in some areas. Like maybe having the money or having the exposure to certain things in life. But I'm not lacking when it comes to like, God gave me something special and I got to figure out what to do with this. And that's something special was just always my brain and my and my ability to like lead with love wherever I go. I got a question real quick because I see the things that your dad sacrificed and did so that you could have a better, you know, education and life and so forth. Where did he get that job to do that? How was he raised? What do you know how sometimes parents tend to because they didn't have I'm going to do everything in my power to make sure that my son, my children have a better life than what I did. Or maybe he saw his mom or dad did the same for him. So he's doing the same for you. I'm just trying to figure out where is the way did it go? Yeah, you know, in the black community, we often talk about trying to break generational curses. You know what I'm saying? I was blessed that at least two or three generations before me in the black community, we just had we had generational love. And we had, you know, generational structure because where did my daddy get it from? He got it from his mama and his daddy. That's my grandparents. They were married for 66 years. So they had that same structure. Same structure.  My grandma just passed away a couple years ago. Beautiful, beautiful relationship. Like when you grow up, when you grow up around that, they get passed down. Right. So so that's important. I got it. That's important. But also you got a lot of people who like I'm a fan of hip hop before I'm an artist, you know what I mean? I grew up listening to rap my whole life. My daddy listen to jazz music.  But me, straight rap, my whole life, straight rap. With that being said, I know these people's stories. I know about Pusha T saying, yeah, I grew up in a two-parent household as well. Imagine every player's aiming coach right. Master recipes on the stove light. I know about Jada Kiz being like, yeah, I grew up in a two-parent household as well. I'm B.I.G. prodigy, DMX and pun. Killing niggas for fun, nothing ill of the sun. Just cause you grew up with that structure, don't mean, what's Pusha T favorite thing to rap about? Selling cocaine, you know what I mean? It's not a guarantee that just cause you grew up with that structure that you're not going to stray and depart from that. Jada Kiz, I saw his daddy do an interview. His daddy was like, yeah, we had him in private school. You know what I mean? So when I was hearing what he was rapping about, I was like, where you saw all that at? We had him in private school. So sometimes people get in the rap game and they hallucinate.


Hi everyone! My name is Katie. If you're new here, I talk about gap years a lot. And this year has been a year for me to really connect with just like so many people who are on gap years. Like I didn't even know that there were so many people. I just sort of like fell into talking about this topic because I pretty much like by default because of COVID I took a gap year from college right in the middle of it. So it's been definitely a weird year. It's been a really weird year and a bad one. But I love my gap year and it has been a really like eye-opening experience. So I think that's definitely something that I want to address in this video. So I've made a lot of gap year content and you could like go and like read the comments on it. And I'm not gonna like add anybody but the comments are there. And you guys often write in sort of your own experiences and I think it's really cool and I think that maybe people aren't paying attention. They aren't reading the comments. Like they might miss some of those comments or fears that are like other people are having and experiencing. The first two, these really go hand in hand. The two most common fears that I see about a gap year is that you're worried that if you take one, you're worried about being productive on it. Or the flip side of the coin is that you're worried about being too productive that you sort of like miss the experience of a gap year. I can't speak to this uh firsthand because this summer because of COVID like I usually work during summer and I did work some in the summer but I had a lot of free time and the whole time that I had time off I was so panicked about getting a job. Then the second I got a job I started doing YouTube and I threw myself into this channel like so hard that I would work three days a week but then posting two videos a week and like a learning how to do YouTube I would literally work seven days a week like sitting on my computer. I definitely over like exerted myself for a while like throwing myself into a project because I really felt that pressure of like I need to do something with this time. It's taken me a long time to like try and find balance. I think that I'm still kind of trying to find balance. I mean especially because I live at home too so that's like another like added layer of like being productive. I don't know if that makes any sense. It's definitely different than like the productive feeling that I get when like I'm at college. Just because you know you fall into old routines and habits when you're back at home so it's that's definitely like a whole another piece of the puzzle that like can get you know confusing. So talking about worrying about being productive yeah like I get this. I would be worried about this too and to be quite honest I think this year particularly I think if you're on a gap year this year like like I barely got an internship and I can see that if like the perfect situation like didn't sort of work out you know fears about going out like into the world and having a job and being around people like that would have been really really hard this year. I think now the more people are getting vaccinated I can see where some anxieties might be like a little bit more lifted this fall and spring. So I wouldn't be too worried about that like productive feeling because you know if you're vaccinated and people are still following guidelines in the fall and the you know the spring and all that which I think the world will be getting getting better at that point. I wouldn't be too worried about that for this coming year because I think the job market's starting to open up a little bit more and you know we should all be feeling like maybe a little bit more comfortable. I can see you know definitely that if you're not like I completely understand that too. So I guess it's like finding the right type of productiveness is definitely important and to be quite honest like and I've said this in other videos like I really feel like I've gotten a lot out of this year even if I haven't like had it all figured out because I definitely haven't uh have had days where I did absolutely nothing and I have I mean when I say I've spent this year like in this room like I literally mean it like I go here and then I go sit like in an office and I don't see the world so it's been a really a year of like not too many perspectives shifting because when you just do the same thing over and over again like you can get like a little bit hamster reel like you start to go crazy which like I've definitely definitely I'm fine I'm fine I'm good I'm really I'm good that's that's pretty much what I have to say on that so I wouldn't be I wouldn't be afraid of this it's definitely I think there's so many fierce surrounding app ears we're like this one but I think that if you were taking it in 2020 like that was definitely something to worry about but with the world opening back up like I think that you know if you want to volunteer or travel like I think even some travel things are starting to open up or maybe spend you know the beginning of the year working and then the later half of the year not I don't know the second fear or third I guess is that gap years are not very normalized okay I feel like this fear was valid like last year when I was taking it I was like literally no one I know has ever taken a gap year except like maybe two people that like I knew of I mean it's not really that's it's not really that scary once you do it I think now it is very normalized the more people I talk to the more I realize how many students this year are on gap years and it's a lot so I would feel less weird definitely that you're like you know in the minority of people who take a gap year because like the amount of people taking a gap year like I feel like has increased exponentially this year and especially after this literal pandemic year like just do what you want you know don't let the haters get down okay the fourth one so a lot of you guys wrote that so this is like this is kind of complicated because it was written like a bunch of different ways but the sentiment is really the same that you are so done with school like so burned out from online school and all that that you feel like going to college next year will you won't have motivation for school or you won't be able to succeed I see this a lot for that I say I think that's the perfect reason to take a gap year to be quite honest I was feeling that way I am so that person like I don't slow down and I like being busy I like working towards a goal and like having that empty year like scared the crap out of me okay I lost my train of thought but having motivation for school I think you know yourself as a person and I before I took this gap year I was supposed to study abroad so I was like oh that's gonna be like my gap year little did I know that's not funny that's really not funny yeah I was like so burned out and like studying abroad is not really like like it's technically like a year off so I pretty much like if I get to study abroad next year like it'll be like I feel like a gap year do over but I'll still be in school and it'll count so like I don't know but I was like so looking for that like I was burned out and not in a way like I love college and like I I love it but I needed a break from I don't know for me like my career and the future and what I'm gonna do with my life like when I'm in college or like even in high school too like those first couple years of college and then the last couple years of high school combined like that is a lot of pressure for one person to handle and I had so many like existential breakdowns and like I just and I think that like a lot of a lot of people have that like you are definitely not alone like and the more people I talk to like literally everyone's like how am I supposed to know I'm 18 and it's like no you don't have to like but like the but you also have to because like the tuition bills coming so like you have to pick a major I don't know it just it all feels so final and like like in some ways it is but then in other ways like it's nothing matters like it's like everything matters but also nothing matters and I needed like a year off to like process that so little did I know that I was going to be getting this extra bonus year so fun so I think that honestly I wouldn't be afraid of not like that fear of like not having motivation I think that you could be a good candidate for a gap year like I think that some time off can can do anybody some good so okay so the next one is that you will lose your opportunity to go to college um this is definitely one that's like very common like I had a panic about panic moment where I was like I'm gonna be 23 when I graduate college like I'm gonna be so old and like I still panic about that you know what but uh other days I'm like no it's fine like I'm just I'm just living life like I think that we all get on this timeline and uh I think like really it doesn't matter and like I have to keep telling myself that to be quite honest but it's really true I mean it's I I shouldn't be I shouldn't even be having to tell myself that because like it is it's honestly true like there is no timeline for your life like things are just gonna happen when it happens and sometimes the world shuts us down and um so I think that losing your opportunity to go to college most schools can let let you defer a year I would just sort of look into that like technically wise but um no college will be there waiting for you like if you go when you're 19 like all the 18 year olds who are just starting college are gonna be like ah a 19 year old what do we do like you're gonna be fine like you know I mean you're gonna make friends you're gonna get right into it uh it's college it's different so that's what I would say and the last one is that you will lose your academic edge yes this is actually a fear of mine because I have to go back to school next year and I have spent a year not exercising my brain in the slightest haven't done a math problem in a year nope not one time have not I still have both of my science liberal arts credits after this which actually I if I study abroad I won't take it for like a whole another year so let's just hope that I remember like sophomore biology yeah you know what I would say for this one like I think this is gonna this is my metaphor for you today like um your your brain is like a loaf of bread and I feel like once the time you like finish high school like it's baked you know what I mean but like this is no this is getting out of hand yeah I think like you the point of high school is like you learn and like middle school is like you learn how to learn and you sort of get like those baseline skills I don't it's not like you're gonna go to college and like forget how to write a paper like I think that those skills are like solidified you just have to kind of trust that you have it in you to to do it yeah I hope this video helped any of you who are worried about this stuff because like I know I have worried about all of these things um constantly so I'm on a gap year I post new videos Thursday yeah thank you so much for watching and comment down below if you have any other fears that I didn't address and I'll comment back to you so leave those down below okay thanks so much for watching