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In this case, the template comes from cheek swabs, trace evidence, etc. You’ve likely heard of PCR being used to test for paternity or presence at a crime scene. more here: īut where does the template itself come from? That depends. We choose what region to copy by designing short pieces of DNA called PRIMERS to bookend the start & stop of this region (1 per strand) so that a protein called DNA POLYMERASE (DNA Pol) can copy each strand. Polymerase Chain Reaction (PCR) is a way to amplify (make lots of copies of) short stretches of DNA from longer pieces of double-stranded (ds) DNA we call the TEMPLATE. Note, this is an updated form of a past post – I’ve included more practical advice on analyzing sequencing data A technique called colony PCR can quickly tell me if my recipe *likely* got in there, but only sequencing can tell me if there are any typos. But before I try to get cells to make the protein, I want to make sure that the recipe got into the plasmid okay and there aren’t any typos. That plasmid serves as a vector or “vehicle” for getting (and keeping) the protein instructions in bacterial cells.
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Basically, there’s a me-altered version of the protein (aka a construct) which I want to express (get cells to make for me), so I took the genetic instructions for that protein and stuck that recipe into a circular piece of DNA called a plasmid. Cloning success! How do I know? Because my DNA sequencing data tells me so! I recently got back the sequencing results for a molecularly-cloned protein construct.