Steps to DNA Cloning

by Jason Amores Sumpter
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in this video, we're going to talk about the steps to DNA cloning. And so recall there are two general steps to DNA cloning and we have them down below label number one and number two. And so the first step in DNA cloning is just making or creating the recombinant DNA molecule. And the second step is going to be transforming the DNA molecule, which recall just means allowing bacteria to uptake the recombinant DNA. And so, if we take a look at our image down below, we can get a better understanding of these two steps involved with DNA cloning. And so, over here on the far left, what we have is the first step to DNA cloning, which is creating the recombinant D n A. And recall that recombinant D N A is a single molecule that contains DNA from two different sources. And so here in this image, what we have is our bacterial plasmid over here and green, and we have over here our gene of interest, which would be from a different species like, for example, a human. And so here this gene of interest is a gene coding for some protein that we're just calling protein X here, Okay. And so what you can see is that in order to create the recombinant DNA molecule, the first thing that we need to do is cut the d n A. And so you can see these little scissors here that we're using as just a symbolic representation, uh, to show you the cutting that must occur in order to create the recombinant DNA molecule. Now in reality, doesn't actually use scissors. Okay? It uses these enzymes called restriction enzymes, which kind of act like little tiny molecular scissors. But we'll talk more about the restriction enzymes later as we move forward in our course. And so the first step is going to be the cut. The d n A. And after the d. N A has been cut. As you see here, the second part here, part one b is going to be to litigate the d n A and litigate the d. N. A is basically like pasting the DNA together, so it's almost like a cut and paste kind of thing. One a. Here is cutting the d. N. A. And one B is litigating the d n A or pacing the DNA together. So you can see we've got these little tiny glue bottles here just, uh, just being used as a symbolic representation of DNA Ligue aces, which are going to be the enzymes that litigate or pace together or seal together these two DNA molecules. And so you've got, uh, these cut fragments over here that are going to be pasted together into a single molecule. So you've got the bacterial plasmid is over here, and the gene of interest coding for protein X is within the same molecule. And so then this, um, recombinant DNA molecule can be used as a vector, a cloning vector to get the gene of interest into the host cell. And that's what we're showing you over here in step number two is transformation of the recombinant DNA. And so, of course, transformation here in this constant contact is just talking about allowing the bacterial cell, which is right here to uptake the external d n A. And so it's going to uptake the external d n A. And be able to obtain that recombinant DNA molecule. And so notice that the scientists way over here is now saying, uh, now that he's got this recombinant DNA molecule within the bacterial hostal. He's saying he can grow this bacteria and allow the bacteria to express as much protein X as he needs. And so, basically, through DNA cloning, uh, we are able to clone the D N A and also create, uh, as much of the gene of interest, the product of the gene of interest that we're trying to create. And so this is something that we're going to continue to talk about more and more these two steps, Uh, step number one, creating recombinant DNA. We'll talk more about that moving forward, and we'll also talk more about transformation of recombinant DNA moving forward as well. And so this is just the introduction here. And so that concludes this introduction to the steps to DNA cloning. And again, we'll be able to get practice applying these concepts and talk more about these concepts as we move forward. So I'll see you all in our next video