Another one, the topic theme, sex and money, who else to demystify this for us other than the Canon blog, Internet entrepreneur and cyber-activist Robert Eli himself is in studio with us. But before you talk to him or see him, VDJ Khalifa, Mambo VB, I see you took us back. Are you trying to make our guest comfortable? Yeah. All right. Okay. So thank you very much, VDJ Khalifa. Mr. Robert Eli. Yes. It's a pleasure to meet you. I'm a big fan. Thank you. The camera is number four. The drill is always introduce yourself if you missed anything in the intro. I think you've finished the introduction. Yes. Yes. All right. So Karibusana. There's so many things people have said about you and around you. Yes. But I want to hear it straight from the Aussie's mouth. Okay. So maybe we can start with your educational background. I'm trained in computer science. Computer science? Yes. I'm trained in forensic. So I think that's the basic of it. But I've always been a techie. You've always been a techie. IT has always been my background. All right. So media. How did media come in? No, just being a loudmouth. Just being a loudmouth. You've always been a loudmouth. Yes. Just being a loudmouth. I'm being pushed to the media by force. It's not my liking. Oh, it's not your liking. Yes.  High school and primary school was easy for you. Yes. It was not so hard. It was not so hard. It was being rebellious, listening to a lot of hip-hop. Listening to a lot of hip-hop. So that's something that you can relate to. All right. So while growing up, did you feel like people didn't understand you? Always. When you're rebellious, you feel like everybody doesn't understand you. So it's not unique to the current generation. And I think it happened to our time a lot. Because when hip-hop was coming up and we were using hip-hop to show our rebellious side, and people didn't understand hip-hop. You remember even Bill Clinton once declared hip-hop a menace to society when he was president. And hip-hop was, you know, hip-hop, drugs, gang culture in the U.S. and so on. I think that's soft side of the American culture. But this was their reality, right? Yes. This was their reality. Relating to that hip-hop, that is their reality. The black Negroes in America. Yes. How did hip-hop relate to you as the youth in Kenya back then? Being black there in the U.S. and also being Negro, being black and in the U.S., facing almost the same challenges our youth in Kenya is facing. Though Kenya might be today from a very extreme background. But you know, they feel oppressed. Not recognized or oppressed. You know, not the opportunities are there, but they're not being treated fairly. So I think the Kenyan youth also feel, even now the Kenyan youth still feel that, you know, the leadership is not recognized. That's why they relate to that kind of music. And so the music was a platform for them to rebel against the leadership and the society. So, you know, we rebel against the father. You start from the fathers, the mothers, the parents, you know, to the leaders. The ideas they try to bring to the U.S. Yes, yes. You remember, we had the Kuflani Maumau, Olamashaka. Yes. Only one idea. If you never knew Robert Eliza, a fan of hip-hop music, you know it now. Who is a hip-hop artist that you can listen to right now, of the new generation? Of the new generation, I don't know. I don't know. I don't know. Maybe Drake, maybe... J. Cole? J. Cole. Not so much. But you know, I think now also trying to understand the kind of challenges we are facing now as a society. You remember now, just yesterday, the women were protesting against the killing of women. But you know, there's also far much killing of men, which nobody recognizes. If you walk outside this studio, most of the people you find idle, 99% of them are men. And you know, some of them, most of them walk at night. They are killed. They are killed.   Nobody treats the killing of a man as urgently as they treat the killing of women. So we should fix the society in general. Yes, we should fix the society. I think we should fix... And when you call it femicide, you kind of leave the boy child out of the war. Out of the war. So the boy child feels that it's me against them. Yes. Which it shouldn't be. You know, you're not winning the war. So it shouldn't be... Let's stop killing everybody. Let's stop killing everybody. Let's not just fight the killing of women or the killing of men. So that the men should protect the women. Yes. That is how the traditional African society was set up. The men took care of the women. Yes, yes. And it's sad that they're moving away from that. Yeah. Very sad. Moving certainly, training in computer science. Yes. Then the internet happened. Yeah, the internet happened. Let's say to you guys. We started, we started, we started with the...  We started from the groups, the young groups, you know. We started from the forums, the list. The chat rooms. The Maschada chat rooms. The list serves, you know, the Kenya forums and so on. Then Google groups happened. Which was more... Which was like, yeah, all groups, but better. In handling. Yes, it was an improvement. Then the web, what they called the Web 2.0 came. You know, now the interactive web. Because, you know, the previous web was one-sided.  Even if you published a website, you didn't have to make it interactive. So the Web 2.0 happened and now it has to be interactive. Now even social media came in, you know, the Facebook and the Yahoo... And the Twitter and the Google chat rooms and so on. So I think it happened and it changed the game completely. Now it removed, it democratized the media. It removed the power from the traditional media. From the big media houses. The big media houses. The mainstream media to now to the common man. Now the most powerful man is not a blogger, not a journalist, but is a person carrying a smartphone. Yes. Can you document something? Yes. Share it with 6 billion people. Somewhere where a journalist was not present, a blogger was not present, somebody just recognized that this thing is uniquely happening. It takes a shot. It needs to be seen. Yes. And we have seen revolutions happen. Yes, yes, yes. We have seen revolutions happening, you know, started by, you know, the Arab Spring and others. Where you see people start from the grassroots and, you know, they document their struggles and it comes and the mainstream media picks up and it blows over the world. I think that's the beauty of the social media. That is the beauty of social media. All right. The revolution in this information age is happening so fast. Yes. It's faster than the previous revolutions. The other ones took a longer time. Now things are happening so fast. Before you know it, we have the virtual reality. Before you know it, we have the AI. Yes. Are you scared? I'm never scared of technology. Because, you know, what's happening with technologies, technology always will frog, you know, our own thought process and understanding of it. You know, nobody understood social media for what it is. We didn't understand that it could have influenced elections. You know, influence of people. Exactly. You know, to decide even sometimes to force people to elect the wrong person. All right. Subconsciously. Subconsciously. Subconsciously. Yes. You know, we have a community in this country which the government is really trying to fight. We are being forced to register with the Department of Defense like the Army and the Navy, the DOD. It's not a military thing. Why should we register civilian thing with the military establishment in this country? So people are scared of things they don't know. Yes. You remember President Moy. He refused to allow people to have fax machines because they believe that fax machines will come. People can use it to overthrow him. You know, you used to have to go through fax machines to be vetted to own a fax machine. All right. Now they are obsolete. This goes back as Africa. Yes, they are obsolete. You remember, I worked in Congo. And my Buddha says, he didn't believe in building roads. He believed that if you build roads, people will drive on them. So you find airstrips everywhere, but there are no roads. Wow. Because he believed that people will drive on the roads and overthrow him. But remember, people work for Muvira. 1200 kilometers away and overthrow him in Kinshasa. So we should... Change the inevitable. No, we should impress technology, understand it, mold it to what can better our society. If you are scared of technology, technology will run over you. There's nothing you can do with the law. And remember what happened with Netflix because you didn't understand it. Yes, yes. And a lot of things have happened. Another thing I've always wanted to ask somebody like you who uses Twitter's platform, do you think the people who designed Twitter expected it to be used in the ways it's being used right now? I don't think everybody understood it. You remember one of the famous quotes, I think it's from Bill Gates, he said that nobody would need so many bytes to hold his data. And now people are terabytes. People are pentabytes. People are going crazy over bytes. People who designed technology would never understand it. Remember that even the person who designed the AK47 said that he really regrets that people can use it to kill. He designed it for something else. Yes, he designed it for something else. So the user always has the cause to play in what? So this is where the regulatory part comes, the government, the society. Because not only the government can do the regulation, the society. Because the government can try to do the regulation depending on the liking of the person in power. But the society wants something else. Like the fax machine then. Like the telephone then. You want the telephone but Mui says no, but people would want to use the phone for their own other means. So I think it's a society to decide what is good. Like now the society is deciding what can we use Facebook for. Should we allow it to be used? You remember the conversations we had I think two, three days ago about the Nancy philosophy video. Which was fake. And it was going viral in the US forums. And people were asking why is Facebook allowing this? But there's no law which was disallowing that thing from being shared. Yes, and there's only so much Zookabah. Yes, so it's the society to decide can you vote on this? Do you want it to be on the platforms? That's why we have referendums. Even on Facebook and on YouTube and Twitter we have the likes and dislikes. We have the upvote and the downvote. So people are subject to their own program.  I think there's no law which can fit. You remember when the EU enacted this privacy bill the GDPR bill. But it is not applicable in sub societies. So you know you can apply the EU law in the EU areas but not in Africa not in the US. So laws are designed to fit the society to itself? Yes, the society needs to decide on what's good for it. Wonderful. As a blogger there are stories that go around. For example in the music business if I'm about to drop something big like an album. I'll get my blogger there. I'll get a mainstream media person there. I'll get my social media person. I'll get my team. The people are going to make sure my stuff is fed to the people. People claim that the same happens in politics. Elections is like an album. So they're going to get the mainstream media guys that they trust to push their gender. They're going to get the print media guys to push their gender. They're bloggers as well. So do you think Kenyan bloggers are participating in this system that we are trying to find first? I think Kenyan bloggers are participating more than anybody else. I think even more than the mainstream media because you remember this is the first government that had a state house blogger. And he changed things. Denis Itumbi was the secretary in the ministry of information at high-to-state house. But you remember because of his work in the run-up to the elections and the re-election of the president, the deputy president. So I think the Kenyan bloggers are really changing the world. And even the last election most of the people were at risk for the bloggers, especially those based at the counties. Because the contest for elective post became so bad that some of the bloggers were being killed, some of them were being kidnapped and tortured and so on and so forth. So I think even today we are in Nyeri also we are supposed to be barring a blogger there. And these things happen, you know. So the bloggers are at the center of something. Yes and they're influencing the influencing conversations, influencing what the brands, what the people want to buy, what the people should listen to. So we agree they have so much power. It's how they use it. It depends on how they use it. Wonderful. We have a video here. I'd like our viewers to watch this video. We are going to throw it back even though it's not Thursday. So let's see the video. Welcome back. Welcome back. All right. We're going to start with the ladies who are doing really well. Come on over here soon. The limit of 60 seconds. But if you do this 20 inch color television set could soon be providing all your evening entertainment. Watching the channel of your choice in the confines of your home could soon be providing all your evening entertainment. Watching the channel of your choice in the confines of your home could soon be a reality. All this yours. If you can prove that you are the smarter one now. I'm going to ask you questions starting now. Who was recently crowned Miss Kenya? Pass. Which is the largest coalition of churches in Kenya? NCCK. Which is the official language of Djibouti? Ethiopian. Which tree branch symbolizes peace? Fun. Which was the highest office served by Joseph Murumbi? Vice president. Which Korean city hosted the 24th Olympic Games? Seoul. Which country? Bonagodana. Which people had Zeus as their chief god? Pass. Which station owns a Shoro FM? KBC. Who is a UN director for Habitat? Pass. Who is behind the theory of survival for the fittest? Charles Darwin. Which building houses the ministry of health headquarters? Afia House. Afia House. You all said that you are going to walk away with something very well, but you have topped it. You win yourself an Ariston gas cooker. Throw away that jico and come on over here. Is this it? This has got to be it. This has got to be it. How do you feel? I love it. You love it? You'll be loving it even more when you get to make those tasty dishes. Who are you going to invite for your very first meal? All my friends in college. All your friends in college. And they're saying to you, he's got it going on, haven't you? Yes. Now you look at them over there. That's the camera. And say to them, I've really got it going on. I really got it going on. This is a good place. Now you know. If you second guest is IQ or is what this? Now you know. So we are Robert Elias studio blogger and an internet entrepreneur. So you won a gas cooker. Yes. All right. Ariston. Then Ariston was the thing. What's the thing? Ariston gas cooker. I don't know if they still exist. I think they still do. But the brand is not as popular. We have the LG's and the Vone. Things have happened. Yeah, things have happened. All right. Where did you take the yogurt? I took it to my sister's place. Have you ever fixed any meal on this guy? Yes. You did fix something. I did fix. All right. It was very interesting to have it. All right. So was this a breakthrough for you? Yeah, 2001. People and everybody was asking me, you know, it was short. It was because I think they shoot many in a day. Then they used to shoot at the Hilton. They shoot many. Then they shot. Yes, throughout the month. So what happened is that after this, I went to Sharks for Christmas. Then we didn't have TV in Sharks. So people just use and there were no phones. So I didn't even have a mobile phone. So after that, you know, when I came back to Nairobi, I was like, oh, it's him. It's him. He was recognizing you on the street. All right. I like this. I like this. I like this. In fact, I think I really still remember the crew then. The crew that shot this. Yeah. Then David Batia, Joanne Chirchir. I think I still... Charlene Samat was the floor manager. I think she's at K24 or something, I think. Do you have a photographic memory or something? No, no, no. How do you know all this? You know, you just remember the people who gave you the opportunity. Because before the show, we used to go for a meeting, like a boardroom meeting and say, OK, this is what will happen. You guys will come for the show. You need to dress this way. And you are not allowed to dress like now we dress. They're checked because I think the camera then were not so good with different colors. So they were not so stable and different colors on the same person. You needed to have a plain shirt or something like that. So we were briefed on what to wear and what not to wear to make the work easier and how to carry on. And you have reached the opportunity so much. In fact, I need to meet Regina Ray. Regina, if you're watching, Robert Eli wants to throw it back like it's there. Yes, yes. Anyway, our topic of conversation today is fame. Because after fame comes the money, then sex surrounds all this. Is this something you've experienced in your so-called career? I don't know if I've experienced that because I was brought in a very strict family. It was not so Christian. It was Christian, but my father was not so prayerful guy. Though every time we challenged him to go to the church, he would unleash the Bible and say that he has known Christianity longer than us.  And so the things go together and people need money to survive. You are not living, it's a currency of transactions in this area. It's the most important thing sometimes too. All right. So money is a currency of transactions. Sex has also been transformed to some form of currency of transactions. All right. As a person in this field, politicians are the biggest celebrities in Kenya. Yes. It's a sad story. People think Kenyans. Yes. Because you know the animals, they have a lot of loose cash. They have a lot of loose cash to go around. Yes, yes. And we live in a hand-out kind of society. Yes, yes, yes. All right. One would expect sportsmen and musicians and artists to be famous, but in Kenya it's different. So in this field, I don't want you to mention things, but have you got people sending you sex scandals from politicians? No, you have to be a blogger in Kenya. Those are the things you have to get. Anyway, I think being a blogger, especially when you cover a wide variety of topics, I used to be only a tech blogger before you started talking about everything else.  Yes, yes.  So I think you get all those things. You get the sex scandals from not only politicians but also celebs, everybody else. How do you go through these things before you can at least publish or share the stories? I think most of the time we don't even share because there are times we used to share and we found out that relationship is very complicated if you don't understand it because sometimes there were times even when we partnered when we were starting these forums and Google groups and mailing lists type then you find that most of the women who abuse, all these bosses in abusive relationships used to contact us. So we partnered with the Koval Coalition of Violence Against Women to be taking the cases there. To be taking the cases there rather than just show them to the world. Yes, but most of the time after the cases had been resolved they were put in safe houses, they were given counseling and so on. Then they were returned to their families for father counseling and they became a couple again. Then the problem started there, that you know, so you become the problem. So the problem comes back to you because you didn't understand the issue. That is what makes it complex. Yes, it's very complex. So if you're not a professional psychologist or a counselor or psychiatrist, sex cases are very, very complicated. Yes, it's very complicated. You don't even get ourselves into them so much. Alright, there's this notion as well that once you start getting some fame and some clout, DMs, Zinaja, Kuja, what's your image? You know the thing is that people take me online to be very abrasive, aggressive guy, so there's a lot of nonsense, you know, just dismiss. Alright. Yes, so DMs and Zinaja are clean, but you know, it's people with problems. Alright, absolutely. You receive a lot, but most of them is people with problems. Alright, so if you want groupies... Blogging is not for you. No, no, no. If you want groupies, blogging is not for you.  Like God's gift to you. Yes, yes, yes.  That is about to happen. Yes. Thank you very much, Mr. Robert. I understand. We are going to take a break and then we'll be back with the game. Okay. Alright, so yes, we have had, Robert, like please tell them how they can find you on social media. Robert, I lie everywhere. Robert, I lie. The good thing is that mine is secured when I... Alright, you're secured. Yes, yes. So even if they Google Robert, I lie, they'll find you. They'll find you.  Alright. Yeah, you know, for your kids and your kids' social media handles. Already. Yes, yes. The future is safe. Yes, yes, because you know, they might need it and somebody else might take it. And you know, it's now an asset. Some people declare it even an asset in your books over count. Their handle. Yes, because it brings you more money than even employment can. Your handle can make you more money. More money than employment can. Some people, you see, they treat every treat, 50,000 Kenyan shillings, 100,000 Kenyan shillings. So it depends on how you... Okay, listen. If you have a job in Kenya or this and that, secure your handle. Your handle can make you money. Yes. And it depends on what you do with it. Walter Robert Eli himself, a renowned blogger and internet entrepreneur. We'll be back with some more of Social Friday. Don't go nowhere. Don't type that down. Why 254? Imagine. Particularly for the man who doesn't know how to stop the music. So since you don't know how to stop the music, from Ni Gani, he weekend it. I don't know how to stop the music. But I don't know how to stop the music. I don't know how to stop the music. I don't know how to stop the music. If you go to Kereguya and town, you should go to Java, then Kesho to go to the Primeval Bamburi. Yeah, Bamburi. Alright. Kesho, Primeval Bamburi. Yes, Kesho. You can go to Nisya, or... Flight. Or flight. We land. In the next life, I want to be a DJ. Robert, how many DJs do you have? Scandolo, Kibao. Yeah. I used to love music. I almost became a DJ. Four in a row. I became a DJ. Scandolo, Kibao. Because that was our topic of conversation yesterday. But this is a social break now. And the social break, we are going to engage you in a game. Yeah, so from my analysis, Robert, I figured you somehow have a photographic memory. So we are going to play Who's Smarter Now or Lover Again. Oh! Is you ready? Okay, okay. Be sure to find him as Robert Eli on every social media. And I'll be regenerating for today. My name is Mary. So let's take it. The second largest continent in the world. I think it's... What? Asia. Asia. Let's see. Africa. Why? Which is the first? Asia. The first is Asia. Asia has to do the BS. I will see. First is Africa. Let's go. The chairman of AU. Jin Ping left. So who do you have now? The Zuma left. I know it, but I don't know the name now. You don't know the name now. But Zuma was the previous one. The wife to Zuma. Yes. The wife to Zuma. The previous. The previous one. Okay. Let's see the answer. Egyptian president Fatah LCC. The chairman of the AU. African Union. Not the secretary general. You know I'm thinking of the secretary general. Or there's the secretary general and the chairman. Let's move on to the next. The largest religion in Africa is... Really? I think it's Christianity. It might even be Islam. All right. Let's see the answer. Islam. Islam. Why? Wow. I've been to school in Africa. We have the northern Africa. Yes. And then we have samples of Muslim religion. Yeah. Like countries like Tanzania is almost 50-50. Nigeria. Nigeria is very big. Senegal. All right. Hey. Let's go. Let's go. Let's go. Let's go. Let's go. What is the name of the bluebird from Twitter? Twitter bird. That's the answer. Which is that? You know if it's blue? How do you call it? Allure. No. It's almost a song. All right. Or sing. All right. The answer, please. Larry the bird. Larry. Larry is a... All right. Larry is a bird. All right. Let's move on to the next. How many calories do you think you can burn by clicking a mouse button 10 million times? Wow. What? 1,000? 1,000? Let's try 1,000. 10 million times. Why is the answer, please? 1. That's a waste. Thank you. What is the next question? The longest river in the world. The Nile? The Nile? Or the Mississippi? The Nile or the Mississippi? I think it's the Mississippi. The answer, please. The Nile. Yes, we got that one. What is the next question for VDJ Khalifa? The nation. I wish we had one for VDJ Khalifa. We are favorite DJs. We are favorite DJs. Thank you very much for playing with us. Robert, thank you very much for coming. And thank you very much for what you're doing for the community. And I hope you're mentoring some bloggers as well. Yes, yes. We have to mentor them. That is in the process? Yes. All right. We talk a lot to the bloggers. Wherever we travel. And also there are others. I have them in my office. Some you have in your office? Yes, yes. We mentor them. The office, you know, it's called Chinyamaji. Office, let's say, go to Chinyamaji. Exactly. But at least we mentor some. We interact because we don't want to leave it to us. You are growing older, you are having... There are things you can't do. Yeah, and things are changing. Which you could risk when you are in your 20s. When you are single and you are not a parent, there are things you can change. And also there are things you can do now. You can support them. You know, you can mentor them. You can tell them the wrongs and the good and the bad side of blogging and how to protect themselves. So mentorship is very important. Very important. One last thing I'd like you to share with the viewers. People always have questions to ask the government. People have a lot of questions. Recently so many people are asking me about this registration process. People are asking me about so many things. And they don't know how to air their questions. They don't know how to get them out there. What is the process? Use your social media. Use your social media. Just be loud. Be loud. Shout. Shout. You used to shout until they noticed us. Alright. I think if you don't shout, if you rely on robotics, sometimes you grow old, you die today. So the power is in oil. If you have a question, ask. And this is the challenge I'm always giving to not only bloggers and people who want to speak out. Also people looking for employment. You know, the power nowadays is in oil. But you know that it took us a long time to even have a dialogue, internet on our homes. That is, you're watching TV through the internet. Yes, to the most remote places. You're watching what is called video on demand. You don't wait for news. You watch it on YouTube. As soon as you read it. Whenever you go hang out. Kitambo used to go to a club. They stop the music. Then they continue the music. People don't care. They play music. And when you go home, you can watch the news you missed. And you can receive the WhatsApp. Thank you very much, Mr. Robert and Lai. For that matter, be sure to find us on YouTube at Y2P4 channel. If you'd like to have a recap of this conversation, or if you know somebody who would like to know what Robert and Lai said on Y in the morning. I go by the name of Bymosis, or it's Bymoney on the social media platform. Don't drink and tweet. I like to tweet.


QuickBooks Online, chart of accounts. Get ready to start moving on up with QuickBooks Online. We're gonna be using the free QuickBooks Online test drives searching in our online search engine for QuickBooks Online test drive, selecting the option that has Intuit.com and the URL Intuit being the owner of QuickBooks. Support accounting instruction by clicking the link below giving you a free month membership to all of the content on our website broken out by category further broken out by course, each course then organized in a logical, reasonable fashion making it much more easy to find what you need than can be done on a YouTube page. We also include added resources such as Excel practice problems, PDF files and more like QuickBooks backup files when applicable. So once again, click the link below for a free month membership to our website and all the content on it. We're gonna be using the United States version of the software and verifying that we're not a robot, scrolling in a bit by holding down control up on the scroll wheel currently at the 125% on the zoom in, hitting the cog drop down to note that we're in the accountant view as opposed to the business view. We'll try to toggle back and forth between the two views so you can get a look at both of them. We're gonna be duplicating some tabs up top as we do every time, right click it on the tab to duplicate it, right click it on the duplicate a tab to duplicate it again, back to the tab to the middle we wanna go down to the reports on the left, we want to be opening up the balance sheet report as that's thinking even though it's done, I'm gonna go to the tab to the right and go to the reports on the left. This time the income statement, the P and L the profit and loss, I'm gonna close the hamburger otherwise known as the hamburger and do the range changing. The ranges, they are a changing from 010122 tab 123122 tab run it, if you wanna refresh it and I do so I'm gonna go to the tab to the left and then close the hamburger again, scroll up change the range again from 010122 tab 123122 tab run it to refresh it. That's the setup process we've been doing every time back to the first tab. We're now gonna be looking at one of the primary lists that they call the term lists which once again is a QuickBooks term it's probably not a term that you'll hear for normal accounting practices but because it was housed under that lists dropdown we can call it the major, the primary list which is the chart of accounts. Now remember in prior presentations we've been focusing in on entering the transactions into the system remembering our accounting goal which is from a financial perspective in order to create the financial statements and then also from a bookkeeping perspective to facilitate the financial transactions meaning dealing with our customers and our vendors and our employees as easily as possible and that means we have these data input forms which are gonna be set up and we wanna put them together so that the data input processes as easy as possible and the creation of the financial statements is as automated, as automatic as it can be. Now to do that we first need the underlying foundational thing which is the chart of accounts. We have to have a chart of accounts in order to make any financial transactions. Any of these forms that we enter is gonna be impacting at least two accounts from the chart of accounts in order to create the financial statements, the balance sheet and the income statement with the exception of like an estimate form and a purchase order which don't have an actual financial transaction but we still track them internally. Now most of the time when you go work in an accounting department you don't set up the chart of accounts so you don't get the opportunity. You're oftentimes working even in one cycle like a customer cycle or a vendor cycle. Small bookkeepers for example, they get to do everything but usually not as detailed of a level. So if you're working out a company then you might not have as much experience adding and subtracting working with the chart of accounts directly. We will get that experience more in the second half of the course when we start a new company file where you get to set up that foundational stuff, the chart of accounts, the customers, the vendors, the products and the services. But for now we'll just kind of, we'll take a look at those underlined under the hood kind of foundational items that we do when we first set things up. Now you could find those. We looked at them last time under the cog dropdown and then lists where we have, if I go into the lists here, we've got the chart of accounts and then you can also find it in the accountant view by going to the accounting on the left hand side and then the chart of accounts. I'm gonna say see your chart of accounts so you can look under the hood. He's looking under the hood here. So we're gonna go into there and if it was under the business view, dropdown, switch to the business view just to check out where it's located there. It's under the bookkeeping, under the bookkeeping. And then you've got your chart of accounts under the manage items. So once you're in the chart of accounts, the chart of accounts is gonna be something that's provided oftentimes when you first set up the QuickBooks online. And you would think it would be tied into more deeply when you do the setup process of the system. For example, the QuickBooks desktop version I think is actually quite a lot better at setting up the original chart of accounts because you select things such as what type of industry are you in and are you a sole proprietorship or a partnership? And then it gives you, you would think, a customized kind of sample based on that information. But the desktop version gives you a much more customized chart of account. I believe the online version still is just giving like one giant kind of a chart of accounts for no matter what you do on the data input. So just realize that you might have to go in here and do more customization that you might be used to if you're selecting kind of chart of accounts from the desktop version, which might be a bit more customized. So just realize that the general rule with the chart of accounts when you set up a new company file would be that you're gonna get the chart of accounts from QuickBooks automatically. And then when you enter data into the system, I would then say if there's already an account lined up that matches up to the data you're gonna enter into the system, you wanna use it typically unless you wanna change it for some reason because you think there'd be a better name for it. And then if you're going to change the account, then you typically wanna go in here and change the name of the account. In other words, you don't wanna make two accounts, mainly for expenses this happens that are similar in name. So permits and then another name for permits or legal and professional, and you just call them professional instead of legal and professional. So if you want one for professional and breaking out legal or break out accounting, bookkeeping it and lawyer separately, then you don't wanna have two accounts that are similar or else you and you're not using one because then you'll probably post to it and you'll get kind of mixed up. And then after one or two months of data input, you might wanna then go into your chart of accounts and look for those accounts that you have not used at all and either delete them or make them inactive so that you can then hone down your chart of accounts to the accounts that you're actually using. That's the method that I would think most people would basically want to go through with their chart of accounts. Now, if you don't like the layout of the chart of accounts at all, you could basically try to delete or make inactive all of the accounts and we'll do this when we get to the bank feeds section because we'll practice creating a chart of accounts as we enter transactions from the bank feeds and we'll enter all of the accounts as we do the transactions. So we'll get a feel for that and that way you can be a more customized, more customizable, but you have to have some understanding of how the chart of accounts is set up or better understanding to do that. So let's see how the chart of accounts is set up. Normally we think of the chart of accounts being sorted by the type over here, the type of account. Now, when you think about the account type, you're usually just thinking first on its most basic level, the balance sheet on top of the income statement. So this is the detail type. I should be over here on the type. So notice it's sorted by type. So balance sheet on top of the income statement. So if I go back, so if we go on over to our balance sheet and I was to collapse the balance sheet down to its bare bones, down to its essentials, I'm looking assets, liabilities and equity. We in essence have the accounting equation. So that's how you first want to think of the chart of accounts in essence. You have assets, liabilities, equity on top of in essence income and expenses. If I jump on over to the income statement and break this down to its bare bones, you've got income and then you've got a couple categories cost to good sold, but basically expenses, income and expenses. So assets, liabilities, income and expenses. If I go to the chart of accounts, you're basically thinking we're sorting by account type, balance sheet on top of income statement, assets, liabilities, equity, which is the balance sheet, and then income and expenses. We'll talk more about how the accounting equation fits with the income statement and so on later. But then within those categories, you've got these kind of subcategories. And these are interesting to note in terms of what they're gonna do to the financial statements. How are they gonna be reported on them and why we have some of the subcategories and how they might differ to what you might see in financial reporting. So let's just get a feel for that. The first one is the bank accounts. So I'm gonna open these up and just give a quick reference. We'll talk more about the balance sheet later, but notice you've got assets, which isn't an account type, but you still have the triangle because the triangle represents the full account category of assets for financial statement reporting. You've got current assets, also has a triangle, but isn't an account type over here. You have other current assets, but not just current assets because for financial reporting, we have current assets. But then you've got bank accounts. Now bank accounts is not what you would see normally on financial reporting. You would see cash and cash equivalents, typically. But here we want to call them bank accounts to indicate the fact that these are gonna be accounts that are connected to possibly bank feeds. They have a special usage, and therefore they have a different account type. This triangle right here is driven by the account type. So you can see we have the account type, and then these two are under that account type. So that's a little different than financial reporting purposes. Accounts receivable also has a triangle because it's a different account type. The reason it's different than just current assets is because it has a special need of a subledger relating to breaking out the receivables by customer. So it's a little funny to have a sub account and the account underneath it, but it's driven by the fact that we have an accounts receivable account type in and of itself. Everything else that doesn't fit in that special category or that doesn't have a special category for other current assets is in other current assets, which is our next account type, as we could see here. And then we've got the fixed assets, which are gonna be the property, planted equipment, you might hear them called depreciable assets, buildings, equipment and so on, and the related depreciation that has its own category there. And then we've got our liabilities under the liabilities. That's an overarching category, a financial statement category. That's why it has a triangle. Then we've got current liabilities, which is a financial statement category, but it's not an actual category on the chart of accounts. We have other current out liabilities, but not just current liabilities because they want to make these sub accounts. Once again, accounts payable similar to accounts receivable has a special need to track the payables by who we owe, by vendor, and therefore it has its own account type. That's where the triangle is coming from, accounts payable. And then we've got the credit cards. They have their own special account type because they might be connected to the bank feeds. And that's why you have a triangle here for the credit cards. And then you've got the other current liabilities, which is every other current liability that doesn't have some special need. That triangle is also represented by the account type. Then you've got the long-term liabilities, another category, that triangle represented by the account type. And then you've got the equity accounts. We'll talk more about them in the future, but you can see that's representing another account type down here on the equity. And then you go to the income statement where you have income. And so that's another account type income accounts. Then you have these other triangles in here. Notice that's not because it's a financial account category in terms of a general account category or an account type, but because we made sub accounts, which we'll talk about shortly later. Then you've got the cost of goods sold, that triangle is created by another account type, as we can see here, cost of goods sold. Then you've got your expenses, which is usually your largest category. You've got a bunch of other sub categories under the expenses, which are not account types in of themselves or financial categories, but rather sub account categories. And then way down here, you've got other expenses, which is another account type. That's where that triangle comes from. And you can see the sorting down below. So that's how you generally want to be sorting the chart of accounts. Now note that you can sort the chart of accounts by name, but you typically don't normally do that. Also note, when I sort by account type, it's not sorting by alphabetical order. It's sorting by balance sheet on top of the income statement, and then more specifically, which accounts on the balance sheet on top of which accounts on the income statement, a little bit more detailed. So then you've got your QuickBooks balance. You've got your bank balance. Notice that they have a register here. We're familiar oftentimes with a register for the checking account, which allows us a more simplified data input into the system, possibly used if I'm entering data directly from a bank statement or something. So it's a little easier than to do the full deposit form and check forms or expense forms this way. I'm gonna go back. Also note, however, this is a way you can get to a register for every balance sheet account. And so if you have a simple transaction, that's not normal, meaning it's not like something you use a form for, such as breaking out between short-term and long-term of a note payable, I could use the register instead of a journal entry. I could still enter a journal entry here, but use the register format to do that. So if there's only two accounts that are impacted, that could be useful, especially if you just kind of want to avoid the debits and credits, because now you can think of this account's gonna be going up or down, and then you can just have the other side go into the other side instead of thinking about debits and credits, for example. Okay, and then we have the batch system up top. So if I select multiple items, I can make them inactive at one time. Inactive meaning that, for example, if you wanted to delete the accounts, you might not be able to do that given the fact that the accounts could have been in use in the past and we don't want to delete accounts that were in use. What we want to do is make them inactive so that they don't show up in the future. So they don't show up in here, which can make your data input very quite long. And if I hit the plus button and do a data input form, for example, notice your chart of accounts is gonna be populating in this area. So you'd like to have it as streamlined as possible. You don't want to have 1,000 accounts in here if you don't need to, so that you can make your data input as easy as possible. You can do that by making the ones you're not using inactive. Closing this back out, I'm gonna say, okay, you can search for a chart of account here as well. And then over here, you can also use this pencil to do editing a little bit more quickly. This might be most useful when you first set up the chart of accounts and say you like the chart of accounts that they have, but you wanna tweak the names to be more customized to your names. This might be a little bit faster way than just going and editing each one of them individually. And so then you've got the printing. You can also hit the cog here and you can change, for example, I think I removed the detail type. You can add the detail type back in or make any changes you want to make the look a little bit nicer. The most important thing here, I believe, is the include inactive items. Inactive means we wanted to get rid of them. We wanted to kind of delete them for future use so we don't see them popping up all the time. But if I wanna go back in and see those inactive items, I have to turn this on so that I can see them. You can also increase the amount that you see on one page up to 300 here as well. So those are gonna be those items. If I was going to run a report then, I can, or if I was gonna make a new item, I can make the new item up top into the detail. So here's your bank account. Here's the account type that we would set up. That would be the main thing that you wanna make sure you get right. You've got your sub account, which is not as important. So don't worry about that too much. You wanna get the name that you want here. You might not need a description. The sub account is where you make that other triangle and we'll see that shortly as we go through the chart of accounts here. Let's just go through them real quick now. So I'm gonna, we got the checking accounts. Those are gonna be having a special need because they possibly have the capacity to connect to the banks. That's why they have their own kind of space up top and aren't just other current assets. You've got your accounts receivable, which oftentimes is required to be there. You might not be able to delete it or possibly even make it inactive because QuickBooks wants an account that ties to the sub ledger if you're gonna have receivables. That's why it has its own account type. And then everything else that's basically in other current assets that doesn't have a special need is then within here, you've got your inventory assets. Now note that when you set up your chart of accounts with QuickBooks online, they usually give you this huge chart of accounts which might include things you don't need and you might wanna go through there and basically remove them. We'll talk more about that in the second half of the course and with the bank feeds or in other courses of bank feeds. And when we set up a new account, but if you don't have inventory, maybe you can remove the inventory asset. Prepaid expenses is another one that's an accrual account. If you're on a cash-based system, maybe you don't want it there or maybe you want a more specific prepayment such as prepaid insurance or something like that, in which case you can change the name or remove it. You've got the uncategorized assets. Anytime it says uncategorized, that's usually what QuickBooks uses to default to a transaction when it needs to make the double entry accounting system in balance but it doesn't know where to put it. Therefore, anytime you see something in uncategorized assets or uncategorized expenses or uncategorized revenue or sales, it's an indication that you need to go in there and fix it, categorize it. And then you've got your undeposited funds, that's a holding account that is gonna be in place so that you can then group your cash into the checking account in the format that makes most sense. Then you've got your fixed assets which include depreciable assets, the fixed assets and the depreciation accounts. Now these, when you first set up QuickBooks online, they often give you a whole bunch of these accounts and you wanna be careful about them because usually you're gonna have to track depreciation related to them and you gotta do it at least on a tax-based system. Oftentimes you're gonna use tax software to do that or get help from your accounting firm to provide the reports. So you might wanna adjust your assets, your fixed assets to line up to the same grouping as your tax software. So that's something I would recommend setting up and understanding before you start using those accounts with your accountant. Account payable has another special need which is to track the payables by a vendor. That's why it has its own account category. The credit cards have their own account category because you can connect them to the bank feeds, therefore they have their own type. And then we've got the other current liabilities, all other accounts that don't have a special category. The, this account represents sales tax. So those would be set up as you set up your sales tax information into the system. Loan payable, other current liabilities here and then would be like a note, for example, current portion and then the long-term portion, notes payable, long-term, typically notes that are gonna be due after a year. And then you got the equity section which is often confusing when you first set up the QuickBooks system because again, I think QuickBooks Online gives you like a whole lot of stuff in the equity section because like I say, they don't really customize as you do the data input towards what you put on the data input, like industry and by what kind of business you're in, they just give you a whole bunch of equity accounts that you could use then. So you wanna pick the ones that are applicable and then possibly make inactive or delete the ones that aren't applicable. So for example, equity represents the owner's claim in essence to the assets of the business as opposed to the third-party liability claims but it'll differ in terms of the names by what kind of business you're in. So first of all, you got opening balance equity which will be there no matter what kind of business you're in but it's not something that you really want to be reporting on. It's just there as a holding account or an account to make or allow QuickBooks to be in balance when it needs to put in like the opening balances. So we'll talk more about that when we do the first setup of a company file, it's kind of a balancing account. So it's a, and then you got the retained earnings. Retained earnings is only a term that you would use if it were a corporation. So if in general, right? So if it was a sole proprietorship, we would call that kind of account the owner's equity account or a capital account. If it was a partnership, you can have multiple kind of capital accounts. So this retained earnings account, whatever you call it, it's special because, and you can tell that because it's a balance sheet account that doesn't have a register. That's how you kind of know that this is the account that the system is closing out into. The closing entries will be done automatically into this account. So you might want to change the name of it but to whatever would be appropriate for that account to be closed out into. If it was a corporation retained earnings would be appropriate. If it was a sole proprietorship, you might want to change it to owner's equity because it would just be one person owning the company file. If it's a partnership, it gets a little bit more confusing because now you've got multiple partners and you have to manage the capital accounts in accordance with the partnership agreement. So you might have one account that it rolls into and then you got to allocate it to the partnership accounts in accordance with partnership agreement, which is kind of annoying, but it's a partnership. So, and then you could have a draws in here and you might have dividends and whatnot. So again, it'll differ depending on the industry. We'll talk more about that when we get to the balance sheet. Then you got your income statement accounts. Usually you don't have a lot of income type statement accounts. This is your income account revenue account, another term that you might call it because you usually do one or two things. You have a lot of expense accounts, less income accounts, but because this is a job cost system, they have a lot of them. Notice down here, this represents that it's a sub account, which means that when I go to the income statement, that's where you've got this little sub arrow down here, which doesn't represent a financial category or a QuickBooks separate account type, but rather a sub account. So that's how you create those sub accounts. So if I scroll all the way down, what happened? I was in here, if I go into this one and edit it, you can see it's a sub account. That's how you create those other sub accounts. Now note that the QuickBooks general chart of accounts tends to kind of like having a lot of sub accounts and it's kind of up to you as to whether many people really like having a lot of sub accounts and many people don't like having a lot of sub accounts. And so you're gonna have to go in through there and customize it. Your cost of goods sold is gonna be there in its own category. If you sell inventory, that's when you would use it. Otherwise, you know, you might not need it. And then your expenses. Now the expenses is gonna be the category with the most stuff in it usually. And it's the area that most differs from company to company. So this is where you're gonna have to spend some time and start to think if you're setting up a new company file, what kind of expense accounts do you want and you could go through here and see the kind of expense accounts that the online system sets up in general often has a lot of sub accounts, which could be good, could be bad, you might go through it and say, hey, I don't want sub accounts for like legal and professional possibly. Maybe you just want an accounting and a lawyer account and have them separate and not be sub accounts. So maybe you want them under the sub accounts. So it really can be customized to your particular preferences. And same for the maintenance here. So maintenance and repairs, I would call it repairs and maintenance, right? Maintenance and repair sounds backwards to me. I don't know why no one says that, dude. It doesn't roll off the tongue that way. But in any case, then you've got your sub accounts in here. Maybe you just want repairs and maintenance. So there's a lot of that sub account stuff and detail in the default chart of accounts that's typically given to you when you first set up a company file. So you gotta go through it. Utilities is the same kind of thing and determine what you actually want. Now, remember how I would do that would typically be that for most people I would recommend you use their default chart of accounts to the extent that you can making adjustments possibly to like the pre-shable assets and stuff upfront and whatnot, but with the expense accounts, try to use what they have in general. And then as you're doing the data input, you wanna think, do I think there's a better account? Would I like to change the name? Maybe I like the account, but I don't like maintenance and repairs. I want it to be repairs and maintenance. Well, then I would go into here and just change the name of it. I would go into here and edit it, change the name. I'm not gonna make another account called repairs and maintenance and have one called maintenance and repaired because then it's likely that I'm gonna post stuff to both of those and kind of mess things up. So, or if I do that, I wanna make sure I delete the other one, right? And then if there's no account at all that relates to what I want because I have a special account that I want in my industry, then I would add the new account as we go. And then after doing like a month or two at least of data input, then you can go back in here and look at all of these accounts and see which ones are you not using, which ones are inactive and then make them possibly inactive so that the data input in the future will be easier. And you can also then go into these items and say which of these items do I like being reported over here in the income statement as sub accounts and which of these maybe are too tedious to be sub accounts and just adjust the sub accounts. And you can do that easily like if this was a sub account, if I had something posted to it and I edit it and I just remove the sub account, not a problem.  You can remove the sub account and then over here it'll show the difference so you should be okay to do that. We'll practice some of that stuff more in the second half of the course when we do data input or another course or in the second half of the course and when we do bank feeds we'll actually create, we'll basically delete most of the accounts that we can and we'll practice actually creating this whole thing, adding new accounts, customizing the accounts as we go as we enter the data with the use of the bank feed. So that'll be exciting.


My name is Jake. I'm with Compose. And today I'm going to show you how to migrate your Heroku app over to IBM's cloud HA PostgreSQL. Before we talk about how to do that, I want to talk about how we actually attain high availability. And the way we do that is with an app we created called Governor. Governor sits on top of SCD, which is our distribution coordinator. SCD is connected to two PostgreSQL nodes in leader follower format. SCD is the one that's maintaining its leader in follower format. And we use Wall Stream replication to keep the nodes in sync. In front of the two nodes, we have an HA proxy load balancer. This load balancer is connected to both nodes and to the governor. So it knows which one is the leader and which one is the follower. If any changes happen to the nodes, the proxy will hold the request until the new leader is shown. So your app won't have any idea that a failover just happened. That's how we keep you with zero downtime. Your app actually connects to the HA proxy node. And so we released this into open source a while ago. And it's been forked a couple of times. And one of the newest forks that came out is Petroni. Petroni adds to governor by allowing you to use ZooKeeper and Console as your distribution coordinator. And it's very stable and quite good. Now we actually use Petroni for all our PG deployments. So that's how we actually attain high availability. So let me show you the demo that we created to migrate over. This demo that I created is an artificial intelligence that's totally real and not made up at all. And what this intelligence does is it will take an image and stick it into one of four Star Trek captain posters. So it takes your image and puts it in there. So let's take a look at what this looks like. So I'll go ahead and upload an image here. Bam. That's Picard as me, obviously. I'm more of a Cisco guy, but this one's not that bad. That one is way better. I'm going to keep that one as my wallpaper, actually. And then obviously we have the other two captains, Kirk. I'm not that fond of. OK, I do like the hairline, though. It's a lot nicer. But then we go to Janeway and clearly the AI needs tweaks. So that's our app. Luckily, we don't have to do anything about the tweak in the AI for what we want to show you today. So for today, we're going to migrate over the data from production, captainly, over to compose cloud, PostgreSQL. So the way we're going to do that is first thing we're going to do is pull a backup from Heroku and capture that down. So go ahead and capture that. And then we'll download it. Hopefully it doesn't take too long. The awesome images that I showed you before? No, that's totally real. It was made by an artificial intelligence, for sure. And not by one of our awesome designers. All right, so now we'll pull the download here real quick. And so now we got the export downloaded. And I'm going to take you over to our Postgre configuration details page. Let me get there real quick. So here's the details page for the Postgre deployment that we set up under IBM Cloud. Here's the connection string for you. One thing I would like to point out, our deployments come with the entire server. So you can put as many databases as you want to on it. We load all the PG deployments with a default database called compose. This has a couple of things in it, like your admin and some other things in it. And then I've already created the captainly staging and the captainly production databases. So we'll go ahead and use PG restore to restore that. So right here, we have the standard PG restore, a couple of options for no owners. Here's the host from the connection string. All of our deployments use non-standard ports for security reasons. So you're going to need to use the minus P option. Then you'll have the admin as the username. And then we're going to import into captainly staging and the dump that we want to import into. Let me go ahead and copy and paste the password, because it's going to ask me for that. Cool. So now that's importing. One thing we're going to need to do on staging is we're going to need to drop the Heroku Postgre add-on. The reason for this is because the database underscore URL config variable is automatically attached to any add-on that uses it. So you can't actually reconfigure it with the add-on. So we have to remove the add-on. And we'll go ahead and remove that real quick. Cool. Now let's double check that. All right, so the database URL is gone. Now we can go ahead and set the database URL. And everyone can see how badly I type. And we'll use basically the exact same connection string, except we're going to change this last part to captainly staging. And let's see here. All right, still processing the data. Almost done. Cool, we're on the sequence. So now we can go ahead and set this. I'm going to do set, not st. Cool, so let's double check that. All right, so the database is already set. So let's also go into the console and make sure the connection is up and running. Cool, imports done. Perfect. So now we got data. Everything is connected. So last check, we're going to go to captainly staging. And refresh, make sure we get no errors. Perfect. So staging, our staging test is done. So now all we have to do is pull the stuff into production. That's going to be pretty easy since we already did staging. All we have to do is change this part right here to production. And let that import, oops, got to get password. And we'll do the exact same thing with the captainly production Hiroku instance. Cool, let's double check that. All right, database URL is gone. Now we can set it. And again, we're going to basically use the exact same thing we did before, except we're going to change minus r to production. And we're going to change this to production. Perfect. Now let's go ahead and run the console. How far have we gotten here? So now let's double check with the console. Perfect. So now we've gotten production pulled over. Now let's just double check one more time in the app. And that's it. We just migrated all the data over to the new deployment. That's all we had to do, without downtime to which app. Right. You have to do the config variable switch. So that's going to give you some downtime. But filling up the databases, you don't have to switch over at that point. So you can just wait until everything's done. And then switch over. I don't know how fast your big Heroku app will switch over all the config variables to all the servers. It should be zero downtime from what I remember of Heroku. But I can't. OK, yeah. So you can, I think you can sync one of our deployments into another deployment. I have to double check on that. Because the way we set up the configuration, I don't know if that will actually work that well. Because one will be a leader, one will be a follower.  So I'm not too sure about that. But hit me up at the booth. And I can ask one of the tech guys when we get there. OK, yeah. So this is done. And so you see up here where it says search. Nothing, there's nothing really to search right now in this app. But let's say I wanted to add a search. If it was super simple, I would just scan across the Postgre tables. But if it's anything more difficult than that, I'm probably going to want my own indexer. And you can come into Cloud here and just create a resource, and let's say Elasticsearch. And we'll put in US South, latest preferred. And then we just click Create. I'm not going to click Create right now, because I don't want to provision another Elasticsearch. But you would just click Create, and then it would look something like this once it was done. And this would be the overview of your Elasticsearch deployment, which would have the connection strings for you to put into ES underscore URL or Elasticsearch URL, whatever the config variable is for your app. And yeah. So that's basically all there is to migrating your app over. That's it. So if any of you all have any questions, let me know. Yeah, that's it for this. So we tried to keep it really short since it was sponsored. And we didn't want to just sell you and pitch everything. So we just decided to show you what it can do. That's it. All right. Thanks, guys.


Is Jesus my imaginary friend? People would try to shame you for believing in Christ, but don't buy in. For I'm not ashamed of the gospel for it is the power of God for salvation to everyone who believes to the Jew first and also the Greek. So should we just give up on these folks? No! God has the power to transform their heart. And I will give you a new heart. I will put a new spirit within you. I will take out your stony stubborn heart and give you a tender, responsive heart. Pray that God would transform each of our hearts in this way. Follow more Christ-centered content.


Yes, okay, so I'll be approaching this differently looking at a cancer exomes specifically at sporadic cancers I'm a PI who works on melanoma genetics, but this could be applied to other cancer types So it's well accepted that cancer is a genetic disease one of the best examples for this is called a rectal cancer Where you have a development of from a normal epithelial cell all the way to a metastasis and these very small phylogical stages are linked to various molecular changes seen right here Now this has become a paradigm for other solid cancers Even though a cancer has been a looked at genetically in depth We still are at the tip of the iceberg and many more alterations are to be found out so I'll start talking about the Platform setup to look for somatic mutations in cancer genomes So there are three main hurdles for this the first is establishing a very high quality tissue bank The next is the sequencing of very large quantities of samples and finally the analysis of the large number of basepers for these mutations These are just two examples of discoveries using unbiased sequencing approaches two kinases That have been found to be highly mutated in different cancer types and that have FDA approved Drugs be wrath has a recently FDA approved drug and so even though there are these hurdles obviously It's worth moving forward and looking for these alterations So I'll start talking about the tumor bank establishment. So we have here An example from breast cancer where you could start off by getting the primary breast tumor Or you could get the metastatic or and you could get the metastatic a brain tumor You could get these as fresh tumors or embedded Some of these tumors could be turned into cell lines Which could be useful as well for a tumor bank and some of these samples could be turned into a xenograft in an immunodeficient mouse Once you have this genomic DNA you could start looking at genome-wide DNA sequencing So what are the advantages and challenges for these various sources? So the advantage for the fresh frozen tumor or an OCT block is Obviously that you have highly reliable data. This is closest to the actual tumor that was resected However, usually you have very limited amounts of DNA from the source It is heterogeneous because you'll have different clones of cells populating that Tumor and it is a labor-intensive extraction It will require pathologist to find out where the tumor lies And then you will get either macro or laser capture micro dissection to get out the tumor cells Paraffin embedded tissue again highly reliable data the challenge again You'll have limited amounts of DNA. It's going to be heterogeneous again You'll need a pathologist to determine where the tumor cells are and with the paraffin embedded tissue You will have DNA quality issues as well because of the fixation procedure and the embedding procedure cell line Of course, we'll have a lot of DNA It's going to be mostly homogeneous because you have a clone of cells that being expanded in tissue culture The extraction is usually very simple There are cases where it's more of a challenge for example in melanoma because you have melanin being produced that could affect PCR reactions so you have to work on making sure that you don't have melanin in that DNA and of course the cell line could be Useful for downstream functional studies But we have to make sure that the alterations are Identified in the cell line do represent the original tumor So it is worth going back to that original fresh tumor to make sure that the alterations are found there as well Xenographed again. We have a lot of DNA. It's homogeneous. The extraction is simple However, again, we might need to make sure that the alterations are similar to original tumor It is expensive to make xenographs more than the other sources And finally you will get mouse DNA contamination and so that would affect your analysis down the line and you need to keep that in mind Okay, the other part of your tumor bank is going to be that you need your normal tissue Remember we're looking for somatic mutations So usually blood would be an excellent source for this DNA However, it's not always available and if you're looking at a leukemia for example, of course It's not going to be relevant So you could go to neighboring tissue But there you have to be careful because you might have some Contaminating tumor cells and now that we're looking at in-depth sequencing You will be able to pick up these contaminating tumor cells in the neighboring supposedly normal cells You want to have as much clinical information as possible About your patients so the date of birth date of death date of diagnosis malignancy stage The location of your primary and metastatic tumors and the various therapies that the patients underwent So this is an example of the tumor bank that we established But obviously this is applicable to any cancer type So in this case we have metastatic tumor DNA the match normal in our case It's blood the OCT block so the original fresh tumor a as well as the matched cell line so alterations in the cell line could always be gone and tested again in the fresh tumor The cell line allows RNA being made a as well as protein lysates and finally clinical information I'd like to point out That we have here three different cohorts It's a really worth while getting several cohorts in order to be able to validate your genetic data Each one of your cohorts have particular bottlenecks And so you want to make sure that filters are being applied to the various Cohorts and not affecting your results. So it really is worth spending the time on getting these additional cohorts So once you have your tumor back There's several quality controls that are worth considering The first is snip detection to make sure that the tumor and your normal tissues are matched the second is To implement an assay to determine that the fraction of the tumors are 75 percent or above That is because if it's below that there's a chance that you will not see loss of heterozygosity or homozygos alterations The assay that we use in our case is looking for melanoma antigens using immunohistochemistry But obviously for different cancers different assays would be applied And then a third quality control is the mutation analysis of known mutated genes to see whether the Percentage that you identify in your tumor bank is similar to what's known in the literature So this is just a schematic of a somatic mutation analysis where you have your patient Sectored you have your normal tissue in some cases you can get a cell line you extract your DNA You sequence the gene of interest You do the same for your normal tissue and then you compare those sequences Now we're looking at a somatic mutation any difference between the normal and the tumor In this case, we're looking at the coding regions as well as flanking 15 base pairs flanking the exons to look for splice site variations as well So if in the past We use candidate approaches now with whole exome and whole genome we can start being much less biased And so as mentioned previously TCGA the cancer genome atlas is applying a whole exome and whole genome sequencing to cancers So this is probably the largest Industry to provide all this data And so they said that there will be at least 3,000 new cancer cases by the end of September 2011 and they are on target So I'll move on to looking at exomes and Touch on how do we choose the genomic DNA source when looking at whole exomes? so We have to consider DNA quality so from fresh tumor I said we have limited DNA from the cell line We are unlimited The fresh tissue is heterogeneous the cell line is homogeneous Now does the cell line recapitulate the tumor biology now different ways to find out whether a cell line would recapitulate tumor biology And so how do you choose it? How do you assess it? In our case a we were fortunate to be able to do whole genome analysis and this led us to To decide which DNA to use for whole exomes So in the whole genome study We compared the fresh tumor to a cell line derived from that fresh tumor as well as blood And we run it through the lumina sequencer and looked for the melanoma somatic variations These are the build statistics and just like a gym and less mentioned in please note the Passing filter depth coverage you see that it's between 34 X and 67 X It's much lower than what's required it for exomes obviously because that does it's more Homogeneous in its coverage and you'll see the coverage that we get for whole exomes And this allows for 92% of the callable genotypes across the entire genome So the bottom line of this study was that 97% of the alterations identified in the coding region Overlapped between the fresh tumor and the cell line However the copy number variations were less concordant compared to the SNVs So based on this we decided to use the low-passage cell line DNA and not the fresh tumor because We were mainly interested in finding the single nucleotide variations And we knew that we needed at least six micrograms of DNA for the whole exome And so we could derive that from the cell line but getting it from the fresh tuned tumor was more of a challenge Also, we knew that we wouldn't have stroma contamination when looking at the exomes. So we decided to do the cell lines So this is the study design. We use the exome capture in this case 14 tumors were done and in parallel the normals were done as well So 28 exomes in this case the sure select a 37 megabase was it was used And so we captured about 20,000 genes And this was done at this so we had elan and then cross-matched applied This was the discovery every one of these studies is followed by a validation and the validation was done in this case using Sanger sequencing So we've seen a couple of these this morning already This is an example from this study where you can see the BRAF gene and its exomes You can see the sure select oligos covering these various exomes and you see the coverage itself And like I said the coverage is in a variable across these various exomes And that's why you need to go very much in depth in your coverage So what quality tests do you need to do once you get your whole exome data? So these are divided into five and we'll start off by looking at the coverage So this is an example of some of the samples that we did like I said the coverage was extremely high It was a minimum of 180 x and this allowed to a percent target region genotype coverage of 90 percent in average and So this is the performance summary where you get 12 gigabases a depth of 180 x and 90 percent coverage with high quality genotypes So the next item that it should be applied once the exomes are derived. It's worth looking for the specificity in your exomes so and this is a Some of the data that we got and how it was transferred it to an Excel file So what can be seen here is the gene name the reference a little the various the variation a little Same for the amino acids and what in addition to this we have the MPG score The gym and Jamie were talking about earlier As well as the coverage in this case We're using an MPG to coverage ratio and what we're seeing is that if you have a ratio of point five and above and This ratio occurs both in the tumor and in the normal sample when you look at Sanger Evaluation most of the time. This is a real somatic mutation. You can rely on this alteration However, if that ratio is below point five in either one of the sample So either in the tumor or in this normal sample the Sanger will not show you that there's a real mutation there A different way of looking at it is this way So if for example, you take about 90 regions and you assess them by Sanger sequencing They're divided like this So the ones that are above the point five ratio will validate and the ones that are below we won't So this allows you to calculate your specificity So you get 97.9 percent coverage rate and 2.4 percent false negative rate Once you apply this you remove 18 percent of the alterations Okay The next quality test is worth doing once you get your whole exon data is to look at the sensitivity so in this case We knew we already did candidate approaches before doing the whole exome And so we already had 47 somatic substitutions that we expected to see in the whole exome Out of those 47 only 38 were present in our whole exome study, which means that we had 81 percent Sensitivity now note that the missed alterations were not because they were not captured They were captured and they were sequenced, but they were simply missed and we're not sure why at this point Okay, so a the next item that's worth considering when looking at you the exome data is the number of somatic mutations that are identified per tumor So graphically seen these are the various samples that were sequenced and you can see the total number of mutations that can be seen There's a variation. There's a certain range But clearly there's one tumor here that has an extremely high number of some of mutations compared to the others And so one needs to identify this there's obviously a biological reason behind it But it's worth noting it and deciding whether this is a relevant sample for this particular study Now, of course, there could be all kinds of artifacts. These are hard to predict, but it's worth looking out for them And so I'll give you a few examples So in one example when we're looking at the data It's worth so we sorted it only not only by the sample but it's worth sorting it also by the chromosome So in this case it was done for patient number nine and there seemed to be out of the ordinary a Large number of somatic mutations on chromosome X. So we look more closely at this and we found that The genotype on chromosome X in nine n so the normal was one a little we knew the patient was a male But his tumor had two alleles in the same precise location as seen in this table So what's happening here is that there's a copy number variation and this is an important item Especially when looking at cancer genomes that you do have copy number variations And so in this case it could be a Y chromosome deletion an X chromosome duplication You could check this for example by fish. We did not but we do know that there's something going on here And so it is worth investigating the underlying reason before including these alterations in patient number nine on chromosome X Okay, so we went through these various quality tests now. Let's get to the actual study So as I said, we have a discovery screen and then we have the validation screen So we select the tumors and then we do the exome capture both of the tumor and the normal samples It is worthwhile doing the normal in parallel rather than the relying for example on DB snip for the various reasons that At this point Jim Jamie and less we're talking about so here's an example where we used a we first got the 14 samples And we found over 300,000 variants versus the reference sequence But once we applied the normal information It went down to about 5,000 so at this point. Yes, it is worth doing the normals as well Here's the validation screens. So then you go up to validate interesting genes in a larger number of samples I put an x-axis is obviously dependent on the number of samples that you have but the more the better Here you need to compare the gene mutation frequency to the expected background and we'll get back to this later in my talk It's very important to do this because this will allow you to find out whether the genes that have mutations of passenger mutations or their candidate cancer genes Again, I'm emphasizing the discovery and the validation screen For budgetary constraints. This is worth doing Exactly like this going for the discovery and then applying validation scaling up of just a subset of the interesting genes Okay, so let's go into the filtering process in more depth So the number of potential somatic mutations are seen here We filtered it through DB snip and went down to 141,000 then we applied the somatic filter. This was a 58,000 now because of the limitations of capture and sequencing some of the alterations that are actually polymorphisms could come up in another normal sample That's not necessarily the match normal and it's worth taking that into account because this is going to be a polymorphism So when you find that alteration another normal again, you filter that out and Then you remove all the non-coding very alterations and you get down to about 5000 alterations so a this is again recapitulated in the This part of the slide We have about 5000 alterations now when you apply the MPG coverage ratio of point five Like I said, we remove about 18 percent. So we have about 4,000 alterations And this is the way the alterations can be divided by missense non-sense splice sites insertions deletions and Synonymous it is important to keep track of these synonymous mutations as well Because then you can find out the ratio of the non-synonymous to synonymous mutations this is important because This ratio a to be is expected to be two to one if there's no selection However, if there's a significant difference from the two to one ratio It suggests that there has been some sort of selection for these mutations And so this particular gene probably has a role to play in the cancer So obviously a track down these synonymous mutations Okay What kind of data do we need to evaluate the drivers and the passengers? So which genes really have a role to play and which are just there and they have a neutral effect So this is a challenging question. It's being dealt with a lot by the field And this is just an attempt to answer the question. It can be answered by statistics by informatics and functional studies So if we look at statistics first as I said the non-synonymous Synonymous ratio if you look at the full exome study it was two to one Suggesting that most of the alterations that were identified are indeed passengers Mutations above background mutation rate So when you're doing your exomes track this down because the background mutation rate is the number of mutations per Megabase of DNA derived from all your exomes Of course, if this is in the literature, it is worth comparing it to your numbers as well Melanoma the number was eleven point four mutations per megabase Okay, the next is to see whether you have any hotspot mutations meaning the same exact Alteration in the exact same amino acid in different samples the likelihood of this happening is extremely low So again, it suggests this would be a driver and then look for the highly mutated genes So searching for the hotspots in our case we found nine novel genes. We're requiring mutations So a obviously this is only 14 samples. So we scaled it up in additional sets And you can see it went up to a very large number of samples And this is a summary of the hotspots and how they scaled up So this is a non-synonymous alteration. It's new it occurs in our sample set six times And in one of the cases it's found in a commercial cell line It is worth including commercial cell lines in your exomes or at least in the validation step Because that allows you a to do functional studies later on and it also allows the rest of the community To actually have a sample that has the alteration that you identified Validated and also do some functional studies So another interesting point here is that we found some synonymous Mutations that scaled up. So it's synonymous mutation that occurred in exactly the same position in three different samples This is interesting because I mean this is not going to affect the protein per se But what is it doing? It is being selected. So it is worth capturing these as well and maybe following up on them This is a schematic of a one of these hotspots And so this is a statistical test to see what's the chance of this happening by chance In order to calculate this you need the background mutation rate that we talked about before and you also obviously take into account The number of samples that were sequenced. So in this case the the likelihood is extremely low So this is being selected for and what is it doing to the a function of this protein? So we know that this protein is a histone acetyltransferase and we know that when it's disrupted in mice It causes lethality and defects in cell cycle progression Bioinformatics obviously is worth applying in this case You can see that the particular alteration is highly conserved in various orthologs So again suggesting it that it is very important If at all possible and it's hard to predict which gene is going to be a found in the exomes But it is worth doing some functional analysis to completely validate and see whether the mutation has an effect on the actual function So this is an example of a functional study that we did a Where we knocked down this particular protein in a cell line that was either wild type or mutant for the protein And we used shRNA and then check the effect of this knockdown on cell growth and apoptosis And so when we knocked it down in cell lines that had the mutation We could see an increase in apoptosis when there was the cells were growing low serum But this did not happen when we knocked down the cell the protein in cells that were wild type for this protein So this is a what's called oncogene addiction So the cells are dependent on this particular mutation. And so when you knock it down, they're more sensitive and they will die So so it is really worth trying to do some functional studies on alterations identified Okay, the other side looking for highly mutated genes So in this case, there were 16 highly mutated genes. Now, what do I mean by highly mutated? We look to see Which genes were mutated in two at least two samples out of the 14 that were exome captured and I wasn't enough to just look at the percentage of Mutations, but we also needed to do a binomial test where you take into account the background mutation rate and the size of the transcript of the particular gene So again when we found these 16 genes, we validated them so we scaled them up in additional samples seen here So here you can see number four when you look at these when you do this p-value calculation Always take the longest transcript size of this particular gene in order to really push this formula to its extreme And then you take into account the background mutation rate as well So here's where you only consider the percentage of the tumor that are being affected when you don't consider that p-value If you sort it by percentage you see these various genes that come up and they're highly mutated But you immediately see that for example Titan is in this list now Titan is the largest gene in the human genome It's a false positive. It's just because it's such a large gene and there's such a large background mutation rate Of course, it's going to be highly mutated However, when you start applying this p-value and you sort your data then you get a completely different table So in this case you get b-rap on the top of the list It's known to be highly mutated and has the lowest p-value and then you get these additional genes Now you do have a positive control here because then when you scale up in additional samples You can see that there's a concordance between the percentage that identified in the larger number of samples Compared to the original 14 samples. So we knew that we're probably in the right direction here And then when you look at the actual genes we went for the second best brief rough Of course is very well characterized a grin to a was never shown to be mutated previously in melanoma At least not highly mutated So if you focus into this then we did what I talked about earlier Look at additional cohorts to see if we can see a similar percentage in additional samples So you see that indeed when you look at these other cohorts The it is mutated there as well and we have the commercial cell lines here like I suggested earlier So it is important to acquiring these additional patient cohorts This is a schematic of the alterations in this particular protein I'm putting it up here because it's worth a looking at the particular alterations once you have a gene of interest in cosmic Now cosmic is a database it developed by the Sanger Institute It summarizes published and unpublished alterations identified in cancers different cancer types and so if you find a particular alteration In your gene of interest in cosmic it is likely to be important And so it's always worth going back to cosmic and in this case We found two alterations exactly in the same position and then we found a third one in cosmic So it's likely that this particular alteration is important Okay, what do we do with the complex exome? So we've been talking about using cell lines, but what happens once you start using the fresh tumor So we don't have much experience with this. We only started looking at it We do know that when you apply the MPG coverage ratio, we can find a Particular mutations in the normal tissue not only in the tumor. So for example, we found the BRAF v600 e Mutation in the normal. So clearly there's a contamination of our normal sample with tumor cells Which is what I said could be could be a problem Also, there's probably going to be heterogeneity. We haven't determined this but that's something to keep in mind So probably when looking at fresh tissues different algorithms different filters will have to be applied Okay, so I'd like to remind that we've been obviously looking at the large number of genes in the genome a lot of their Exomes and so you can look at genes, but also you can start looking at pathways And this was done for example for pancreatic cancer where a 12 core pathways Were identified to be altered in this particular cancer type, which means that if you look at two different patients Even though they have different mutations in different genes these genes still affect the exact same 12 core pathways So it is really worthwhile applying your exome data to pathway analysis and see how What kinds of pathways come up to be significant? Now is it worth delving deeper? So we talked about 14 exomes Once you complete your first set of exomes. How do you know it's worth doing more or are we done? So this is just a graph showing the mutations per megabase identified for different cancer types Now since we focus on melanoma I have these numbers right here derived from different studies and what seems to be the case is that at least for melanoma There's a very large number of mutations per megabase in some cases in order of magnitude higher So that seems it does seem that it's worth doing more exomes just because they're going to be many more passengers And so when you're doing your exomes, it's worth finding out what the mutation per megabase is Comparing it to other sample other cancer types and seeing you know, is it worth delving even deeper? So what are our future challenges? Obviously finding out what are the drivers versus the passengers from all these alterations How do we analyze interpreted all the data? a The functional studies are pretty important But how do we do this in a high throughput fashion and ultimately how do we apply all this data back into the clinic? So once we have the genes we get the pathways, but we could also Integrate all of this further and see how the various pathways Cross-talk and so this is the kind of database. We're hoping to derive a from many many exomes And this point I'd like to acknowledge all of these people in the slide. Thank you