Components of Dideoxy Sequencing

by Jason Amores Sumpter
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in this video, we're going to focus on the specific components that are needed for diet the Oxy sequencing. But we're not going to get into the actual process of diet the oxy sequencing until we get to another video later on. In our course in this video, we're only focusing on the specific components, and so the components that are needed and died. The Oxy sequencing reactions include the following five components that we have labeled down below a through E. And, of course, these labels a through E that you see here correspond with the labels a through E that you see down below in our image. And so the very first component that is needed in a specific diet the Oxy sequencing reaction is, of course, going to be the unknown template D n a. Of interest, whose sequence we do not know, which is why we want to conduct di di oxy sequencing. And so over here, what we have is our template, d n A. And you can see this is the d n a. Here, and we do not know the sequence of the DNA, which is why we want to conduct this sequencing to figure out what is the sequence of this DNA. So that is the first component, then the second component that we are going to need is actually a d n a prelim arrays, which, you might recall, is an enzyme that prelim arises or builds d n A. And so DNA polymerase is the main enzyme that's needed for DNA replication. And so what you'll see here down below is the second component that we need is the DNA preliminaries. And, of course, this is the components of Di di Oxy sequencing. Okay, Now, uh, the third component that is needed is going to be DNA primers, and these DNA primers are going to a kneel to the template strand. And so recall that DNA primers were used in the specific technique that we talked about in our previous lesson videos called proliferates, Chain reaction, or PCR. And so what we'll see moving forward is that Di di Oxy sequencing is actually going to use a special type of PCR, and we'll see that moving forward now, the, uh, DNA primers you can see down below right here. Uh, and there will need to be, of course, uh, to DNA primers. Now, the the fourth component that is going to be needed are all for D n a, uh, deoxyribonucleic nucleotides. And so these are going to be the normal deoxyribonucleic tides that are used in, uh, d n A during normal cellular DNA replication. And so these deoxyribonucleic tides include D A t p d t t p d g t p and D c T p. So this is basically the Adnan timing guanine and cytosine, And then you can see these deoxyribonucleic tides down below. Here and again, these are the normal DNA nucleotides. And then, of course, for Di di oxy sequencing, where the fifth and final component that we're going to need is a small amount of a single die d oxy rebo nucleotide. And so this is the special type of nucleotide the D d a t p s d t t p S d d g t p s and D. D uh C T. P s. And so, in a particular deity oxy sequencing reaction within a single test tube, we would only want to use just one single, uh, nucleotide. And so we would have to separate. Uh, these reactions using different test tubes to use different dia de accion nucleotides. And we'll talk more about this as we move forward in our course. And so recall that these ddn tps these die deoxyribonucleic tides, they are going to terminate the DNA synthesis due to the presence of a three prime hydrogen atoms or groups. And so what you'll notice is down below right here. You can see that these are the die d oxy Revo, uh, DNA nucleotides. And so these are the special chain terminating uh, D d N T. P s diary, Oxy nucleotides. And so these are really the five major components that are needed for Diaby Oxy sequencing. And these are the components that you will see mentioned as we move forward and talk more about the specifics of the process of diet, the Oxy sequencing. And so, for now, this year concludes this lesson, and we'll be able to talk more about this as we move forward.