![]() But basically, the gel electrophoresis will help us separate these DNA fragments based on size and based on charge. And I made a video on gel electrophoresis if you want to refresh, you can watch that video. So, specifically we're gonna do a gel electrophoresis, "electrophoresis" on these DNA fragments. Now Step 2, what do we do? Well, what we're gonna do is we're gonna take all these tiny little DNA fragments and we're gonna run them on the gel. And that will result in lots of these smaller pieces of DNA. We're gonna expose it to enzymes that will basically cleave the DNA in a whole bunch of different parts. So, we got this big strand and we're gonna cut it up. So, we're gonna take this big old strand. So, "take the DNA and cleave it." So, let me draw that out. So Step 1, what we're gonna do is we're gonna take this DNA and we're gonna cleave this. And we'll break it up intoĪ couple of different steps. Now, in order to figure out whether or not Gene A is inside this cup, basically we have to do this process known as a Southern Blot. If it's inside of this long piece of DNA. I'm interested in Gene A and I want to see if Gene A is inside of this cup. And there's just lotsĪnd lots of those DNA and let's imagine that I'm specifically interested in one gene. So it's got just a wholeīunch of DNA inside. So let's imagine that we have a cup and it's filled with DNA. So, a Southern Blot basically allows you to visualize a specific piece of DNA that you're interested in. Restriction sites: specific sequences of nucleotides, which are recognized by restriction enzymes.Gonna be talking about something known as a Southern Blot. Restriction endonucleases: commercially available enzymes that can target and cut a specific region of DNA they are naturally found in bacteria Gene cloning : a biological technique that creates many identical copies of gene (or any other DNA fragment) and directs its replication within a host organism Sugar-phosphate backbone: t he outer support of the ladder, forming strong covalent bonds between monomers of DNA. Plasmid : a circular DNA in bacteria and protozoa that is separate from the chromosomes Recombinant DNA : single DNA molecule that is formed by combining DNA molecules from at least two different origins Each restriction enzyme recognize a specific, short DNA sequence, a restriction site, and cuts each DNA strand from specific points. ![]()
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