 So it's going to be a really nice continuous of the real stuff, because I'm going to talk about innate profiling. And basically it's kind of the 21st version of the 21st century version of the fingerprint. You know, maybe in this pause I can describe what we are doing here, actually we are doing experiment. And we are trying to be using just the library from 10 random people, actually we are exactly here, like 10 of these people. And based on this the library people are trying to predict and tell what's the gender of the person. So how does this work, actually we will describe later. 1982, 20 years old girl was killed in the middle of England. And the point is that after 4 years, a year of the same city, and now 40 years old girl was killed. And the same crimes, everything really looked similar. And then he just found the one guy who reached the bottom, and he suspected as a killer. But there's one problem, there's not enough proof. And there's one issue with him, that he is mentally sick. And he's changing my degree very frequently. So the question is, is reaching the bottom really criminal or not? The second snapshot, DNA paternity testing. This kid, we have a kid, we have two parents. And we have an investigation, we really want to know, is this two parents really parents to this kid? Or there's another minor woman parents to this kid. So basically, the main question across here, how the stories which I just told you, the criminal story, this story related to each other. And the thing is that we're pretty related, because under the hood, there's one method which both stories utilize to really to answer for the two questions. Like question, who is the criminal and the second one, who is the father and who is the mom. But basically, I just want to kind of convince you that this method, that actually finger bleeding was really method which may break through in one century ago. But now, I would say the new version of the finger printing, it's actually DNA finger printing. And this method is really making and have done huge revolution in the field of criminalistic, especially. But the problem with finger printing, DNA finger printing is actually pretty clear. Imagine, we have our finger prints from the crime scene, let's say in some bottle, and you have 10 suspected people. And what should you do? You should just take the finger prints of these people and try to find the emerge, try to find the person who has the very similar finger prints. Okay, it's fine, but how can we do with DNA? What should we do with DNA? To really find the magic in person to person and all of the modern criminals who are actually pretty smart. We're kind of using gloves or something like this. But basically, based on, I don't know, piece of saliva, piece of the blood, can we find who is who? And actually we can. But to really understand how we can do it, we should go from heart to bottom. So, we know that there we have all our body consists of the cells. And inside the cell there's a chromosome, and in chromosomes there's a DNA. But actually, what I'm showing here, this is actually a super nice experiment you have done in a year in university. We actually were taking cells from the check, sorry, and making a suspension of them, and actually making drop of their cells on a piece of glass. And the point is when this cell is crashing against this glass, we're actually exploding. And all things from inside, like spreading around with this glass. And basically, using some special colors, you can see the chromosomes. This is a really chromosome of two different persons. And the question is, do we see any difference between one sample and another sample? I guess it's more hard. Yes, so the point is every person has 22 chromosomes, 22 pairs of chromosomes. One from one, one from another, from father. But there's one special pair, this kind of gender pair. One, in the case of men, it's XY, in the case of a female, it's X-Tis. And these two samples, we can see there's a difference between them, one of them for a man and one for a woman. Okay, it's fine. Now we really understand how to distinguish men and like female and male samples. But the question is, how can we distinguish samples from different females or from different males? So you trust me, we cannot use this method. The chromosomes look really pretty similar, like within females or within males. So we should really go deeper, we should go deeper and we should go on a level of the molecular level. But there's one problem, we really should zoom like six orders. It's really hard to imagine what does it mean, six orders or nine orders. And really to say to you what does it mean, how to compare, how to make a feeling, what does it mean. Imagine this, this is the diameter of this pen is actually, as well as the same as the diameter of DNA. We have kind of special world like this. And the point is, in this imagination world, one meter distance going to be a distance between a center of Earth to the Moon. So it's really super hard to work with such scale. And we should really invest in our technologies to do it. But first of all, we should really understand what DNA is. So DNA is a linear hetero polymer. And what does it mean polymer? Basically polymer is a special type of molecules where we have certain blocks. And these blocks are repeated. In the case of DNA, it's really linear. And the thing is that these blocks can be the same, but can be different. And in the case of DNA, these blocks actually were really different. That's why we named DNA hetero polymer. Because one block can be either adenine, either cytosine, either guadine, either thymine. And for simplicity we just named them A-C-G-T. And the thing is that DNA double stranded. It's super tricky actually, polymer. We have not just one strand, linear strand, we have two strands. And we're really interacting between each other. When you see there's a small interaction between them, and we're really, really weak. The point is if you increase the temperature until 95 degree, which actually we have done anyway. So you're really able to destroy these connections. Okay. The next property is the complementarity rule. Maybe you notice double stranded DNA. And in this double stranded DNA we have just two types of players. A against T and C against G. And nothing else. You will not find the player like A and G. So this is exactly the complementarity rule. Why is it so important? Because it's really dictate how life can multiply yourself. Like how we can replicate ourselves. You just need to kind of destroy yourself, like remove two strands, separate two strands. And based on the complementarity rule, just fill the rest. And as a result, instead of just one DNA, you will have two. So it's exactly the most important property of the life, division. There's another tricky thing. Actually I was ashamed by my first supervisor in the university. I didn't know about this. It's actually super important. So there's a strange direction. You see these sugars. Actually, sorry, it's sugar here. And you see there's a certain direction of this sugar. This actually corner looking all the time on up. And in this case, in this trend, sugar looking down. Actually as a result, people say, okay, let's say this going to be forward direction is going to reverse direction. Perfect. Why it's so important? And there's a small reason for this. There's a machine, actually molecular machine, which doing actually replication of DNA. So first of all, this machine needs certain situations when you have a single strand DNA and small piece here. You know, you rename this piece like a primer, because DNA cannot start from, sorry, DNA cannot start from the empty place, DNA cannot really get some primer. And DNA cannot really find this primer, sitting in this place. And next asking, okay, on this strength, which should we attack standing right now? Okay, T. So I should really catch from the surrounding of the cell catch adenine and insert inside, insert this DNA. And, etc., we see there's a gene, so we should insert, based on the complementary to C, we see here C, and we should insert G in the front. And the actual thing, maybe you noticed, there's actually three bolts here. And in DNA, there's just one ball. And actually, these building blocks were actually pretty clever made. In the same time, we're building blocks, but in the same time, we're butt batteries, we're kind of energetic batteries. So pulling rice is kind of, you know, terrodynamic machine, and it needs some energy to do some stuff. And this is exactly the, this machine is finding the energy. So, yeah, sorry. So, okay, let's come back to our previous question. What's different between people? And actually, maybe you know, there's one key difference, mutations. And actually, I would say this is most important difference between all of us. We have actually, in general, we have one mutation per average, which make difference me and you. One huge genome. And actually, it's not, it's not, except mutation, there's one more difference. And actually, it's actually really important to say about this difference. And this difference is microstatolide. Basically, it just means that in a certain position of genome, we can have either one gene, but either five gene. So somehow it happened that the number of the nucleotides in certain positions were very variable. So it can be either one, in my case, in one chromosome, and five in another chromosome. But in the case of Lyon, for example, this can be six and seven. And actually, the thing is, this is exactly our thing to bring, if you think about mutations and these repeats. But if you come back to the 80s, cool 80s, you will understand that it's much easier to measure the length when measure which nucleotides you have in certain position. And actually, what we're doing here is actually super close to not to the mutations, but it's really working on the principle measuring on the length of this microstatolide. So how we can measure this length back to the 80s? So in order to really measure this, we need actually one important thing. Julie, you're small, show me. Okay, thank you. So this is, it's not dangerous, it's out of the equation. So this is exactly like agarose. Maybe some of you know, so what is it? Like, actually, I think in the kitchen, in my mom kitchen, I for sure can find agarose because she's using it for jelly and super nice things. But basically, on molecular level, agarose looks like a jungle. There's a lot of trees. And if you are small mouse, if you are small dename, you can really pass through this jungle super fast and super far away. But if you are huge elephant, or huge dename, it's super hard to make several meters, walk forward. And this is actually super applied to the dename thing. It's super jelly. So, just think about the mouse, as I said. Mouse can walk like this, elephant as well. But the point is how a dename can walk. The point is we utilize one super important property of dename that's dename negatively charged. And as a result, we can use this machine. Can I touch it? We can use this machine to make electric fuel. So, because the dename is a negative charge, we can put dename into this gel, into this jungle, and force dename by electric field to move through this jungle. So, as a result, we really can separate short dename and long dename. And actually, this is exactly what we're doing here right now because there's 165 volts and dename walking. I will stop for a second. And you see there's a kind of, you see this color. There's exactly dename going on. Okay, it's not pretty dename. It's actually a special color which going in front of, let's say, single nuclear diet. One nuclear, it's super small mouse, let's say. And it's just showing you the front of the dename. And actually, right now, I can't even experiment because we need higher resolution. We need to give them more time for mouse and elephants to go forward. So, but let's say you want to do it in addition at home. As I said, you can find agarose at home. Don't worry, it's written like homolecular biology can happen. So, what should we do? You should, to make this gel, which you actually touched, you should take water, put the agarose inside, and boil it in microwave, easy. And the second thing, and it's super important, you should add a special colorful compound. This special compound super dangerous. It's actually super toxic. Because this compound interpolate, like biting very strong with dename. So, whatever biting to the dename by default is muta putagena generator, let's say. So, and the thing is that's next to make some sort of shape of this gel, we are putting it in this fancy box. And that's it. So, the next, actually, next step, which you should make. You should actually understand what's polymerase chain reaction is. First of all, when I say polymerase, but when I say chain reaction, what are you thinking about? Like, what's your imagination, how it's working? What kind of, what's, ah? New hairball. Yeah, actually, Julie, my girlfriend, she told me, domino effect, yeah. But in my brain, it was a, sorry, nuclear explosion. And the, but actually, the thing is that you can do nuclear explosion, let's say, in a safe way, in a lab. You just have to add this, you remember, polymerase, dename polymerase. And you should have six components. First of all, it's really important. You should have passion. It's, we really repeated this experiment numerous amount of time. And the real story, Julie, for several days, for the results, deep, et cetera. So, it was really a lot of work. And it's not working from the first time. And you should really optimize a lot. The one piece problem is actually it's a lot of from the people. Because it can be different. Let's look like this. Sometimes it's super hard to clean it and et cetera. So, as I said, you need to have some sample, biological sample, actually, it can be not just saliva, it can be blood, it can be piece of skin, it can be nail, and whatever, you can imagine. Yeah, hair, of course. And one of the most important thing why it's named polymerase chain reaction, you should add the poly, sorry, not just polymerase, it should add spicy hot polymerase. What do I mean spicy hot? It just means that this polymerase should really be stable in the boiling water. In 95 degrees, you know, if you put X in 95 degrees for a long period of time, you will have denaturation of the F. But if you put the spicy hot polymerase, it should be stable. It should be functional all the time. You may ask, where did you find this polymerase, spicy hot polymerase? No, we didn't find this in Mexico. We actually found the thing in Yellowstone Park in the United States. There's a special bacterias, I think we're living in the United States, we're living in a hot spring, and a super red stable, like to live in this temperature. Okay, and one more thing, you need building blocks. Remember, I told you, we need all four building blocks for polymerase to work. One more thing, and actually we will understand later, we need a liquid nucleotide. How did you find this liquid nucleotide? It's actually now a question. We went to FBI website, and we found specific liquid nucleotides which used for the criminals. We really ordered several liquid nucleotides using money from the government. Let's not talk about this. So, and the last thing, so super watered, you should have cool and hot green machine. What are you cool and hot? It's just this machine, it's kind of the refrigerator, but at the same time it's a boiler. So, it's kind of a machine which can change temperature super fast. And actually, next you will understand why it's so hot. So, we took all these six things together, especially passions, and put in one tube. And actually, if you imagine this tube, we have just a few DNAs because usually criminals are living in just one air, not like all air from the head. And our lovely spicy hot boiling rice, which amount of nucleotides, if we phosphate, and the oligonucleotides. And everything going on super peaceful in 24 degrees. Everything fine. But we may increase temperature until 95 degrees. So, we basically, we almost boiling water. But most of the things which are going on here, they actually destroy connection between DNA, between two strands. You can separate them completely. This was connection between two strands, now there's another. And the thing is, next we're decreasing temperature. But not until 24 degrees, we're decreasing until 72. The point is that there's a bacterias from Yellowstone Park were living in 72 degrees and were feeling really pretty comfortable in 72 degrees. And this polymerase evolutionally was made in such a way that this polymerase perfectly working in 72 degrees. But not 73, not 71, perfectly adjusting 72 degrees. We're decreasing this temperature until 72 degrees. This oligonucleotides, which we found in the FBI list, in the FBI website, we kind of found the region, and the region on DNA, or where we complemented two. And we designed this, sorry, FBI, designed this oligonucleotides in such a way that's why kind of we're sitting just before repeat region and just after repeat region. And actually it's super important because this oligonucleotides will allow us to multiply, to amplify especially specifically this region just before and just like, just between these two oligonucleotides. So we increase until 72 degrees. Oligonucleotides found complementarity there. And the spicy hot polymerase found this superfinding combination. And it stopped working. We give some time, give actually a few dozen seconds. And polymerase hardly growing and work, do a job like finding nucleotides and searching, finding out that it's searching, moving forward. And the point is, and now we kind of closing our loop, we're increasing again temperature until 95 degree. And now just make a impression. This is what was previous 95 degree. We have seven pieces of DNA here. But now, when we increase the second time 95 degrees, we have already two times more DNA inside of the tube. And the thing is that's actually, there's a reason why we need this reaction as a chain reaction. Because after every step of increasing 95 degrees, decreasing 72 degrees, increasing the 95, we just increasing number of DNA of this specific region even two times. And the point is we're doing this 30 cycles. So basically if you just have one DNA in the tube, I don't know, it's actually happened sometimes. For example, if you know about the other project, sometimes it's really very hard to find any DNA in the bones. But if you're speaking about criminals, criminals can have just one hair here or one cell. And after 30 cycles, just one DNA will give us two in the power of 30 products. So imagine we are really increasing amount drastically. And the thing is that we're increasing amount of DNA so much that we already will be able to see this by body. You remember this interpolating color like we used to prepare gel? Not this gel which you are actually touching right now. Gel which is sitting here. This interpolating compound just highlighted the DNA where there's a huge amount of DNA. And actually the electrophoresis allows us to separate these pieces. Okay, let's go through the example DNA paternity test. Imagine father, well, has like in certain position on genome have one repeat of G and three repeat of G on the navochromosome because we're not one father. So and the female has two repeats on one chromosome and four repeats on the navochromosome. And now if you take the samples from them and put them in a gel like after all these VCR things after cleaning, blah, blah, blah, you will see like such picture. This is positive charge DNA started from the up and down. And as a result, a small mice mouse pass really far away like close to the positive charge to the anode. And the second one is two lengths like two nucleotides, three nucleotides and four nucleotides. And the point is let's imagine we have a kid. We kind of, we give one chromosome father gave kid one chromosome, one gave a navochromosome. And as a result, we can have just one of the fourth combination. We can have either combination in the case of kid one, two, one, four, either three, two, three, four. So in this case, I just show the example of the, actually as you see one and four. So if you run this really real situation like this, you will say, hmm, look at this. We kind of seen that this kid can be generated by this parents. This distribution can be generated by this parents. And basically the problem, there's just one small problem that's this one, one and four for kid can generate a two person from the street. It can happen by chance. That's two person from the street have the same repeats. And how we can solve it, this problem? The thing is that this repeat regions, it's not just one repeat region in our genome. There's a multiple. For example, in FBI using 16 regions to detect criminals. And there's actually a lot of this repeat regions in our genome and very highly variable. And to really to increase the robustness of our method, if we measure not just one repeat, we measure 16 to decrease really probability that's just two random people from the street can generate a button for a random kid. And okay, let's come back to the criminal story. The thing is that the first girl was killed in 1982. The second girl was killed in 1986. And but this method of measuring of length of microsatellites was invented in 1984. And actually investigators get to know about this method. And I say, let's try to use this. And what we have done, first of all, two really fast steps. First step, we mentioned the, sorry to say, we measured the sperm from the both girls. And we proved that the person who killed them is the same person. We really measured the length of this satellite and we're shown that's where it's similar. So this is the same person. And next thing, which we have done, you remember this poor guy, Richard Buckland. Actually, we just compared the repeats of the Richard Buckland and the sperm and we're sure that this is completely two different persons. So Richard Buckland, not criminal at all. And next, this was routine actually because you should really find a person who has this pattern of the repeats, this amount of repeats in this chromosome, this amount of repeats in this chromosome. And actually, we kind of really forced population of several cities around this criminal place, forced to give biological material every month. And six months, we're doing this experiment which we're doing right now. And these are the new success. Until the moment when one person, person X came to the police station and said, you know, where's one guy in the city calling pitchfork? He's paying me $200 pounds every month due to the length that I go and give you the biological sample instead of him. So that's why this police station, police people came to this person game and took the biological samples from him and proved that's actually exactly calling pitchfork is the criminal and he killed these two girls. And the thing is, there's a historically very important thing that it was the first, first precedent in the law that one person was kind of moved to the jail and his criminality was improved by DNA testing. Okay, let's come back to our test. In this experiment actually we just took, I should use this computer, sorry. So we just took samples from 10 people in this room. Actually, can you raise the hand? Like, she running for sure. Like, I took the, like 10 samples. And I didn't make actually any paternity test. That's not like this. And actually I would say that if you're going to do, if you're going to do paternity test without informing person, and you're going to do paternity test using his biological material, you'll pay $5,000 in Germany and it's actually in several countries you can go to the jail. So be careful about this. And I was careful, but I measured just actually one thing. I measured just a white person. We just took the, we know that's kind of we had 10 people and actually four of them, I think we were like the girls. And the girls we do not have white, white chromosome and girls have. And we just made a PCR reaction. Were you ready? So, okay, sorry. We just made a PCR reaction against white chromosome. That's it. So it just means that in the case of guys you should have a band. But in the case of girls, you shouldn't. So let's kind of make this camera. So, I'm sorry for this. You see anything? Yeah. So, can you close your eyes? So, okay, I just put 70, okay? So, the thing is that this is UV lamp. You remember this colorful thing? Yeah. There's a colorful thing making some sort of light or some sort of weight when we kind of making like lights in this color, this compound by UV lamp. Actually super strong UV lamp. You should not look at this UV lamp without special defense. Can I do like this? No, you can't. The thing is, I don't know, do you mind if I think correct? Me? Okay. So, oh, I'm sorry. Okay. So, the thing is that, oh, okay, look. Guy, guy, you see this band? Oh, sorry, I'm sorry. Sorry. Yeah, I mentioned this. You know, actually, it's our joke. It's gonna be a lot of pain for the eyes. I don't speak about catenoma or like melanoma, blah, blah. I'm speaking about painful things, eyes. So, the first two persons, first two persons guys, like one guy and we see the light, you see? So, it just means, yeah, yeah, super cool. This guy has a white cross on. Boris, of course I'm here, of course I'm here with him. I didn't, perfect, sorry. Boris also have, Aliona, are you here? Aliona's here, but she never, he can't call her back. She doesn't have a white cross on, as well as Diana. Hey! What's the size of that? Joe, I'm sorry, okay. Alex, okay, Alex also have, like Alex, okay. You have, actually, it's a white cross on, sorry. Okay, I'm sorry, but she made mistakes inside. You yourself, you kind of, you're too late. Now, you can imagine, you really need passion for these things, like, and yeah. Anyway, so Julia also have, she doesn't have, sorry, she doesn't have, you're welcome to the men's club. So, yeah, actually, the last slide, I just want to say, really thank you to Julia. She really supported me a lot in all this, I got enough experience. And I also want to thanks to a huge amount of people who gave us so much love, so much support. And I also want to thank you to all the people who gave us this live, if you question what they're doing, why keep my slides, it's my biological material, you should not sequence it, et cetera. We kind of officially say, we just want to check, do we have live from us or not? So, that's it.