We performed a DNA extraction with the same protocol that we previously followed. We then used this extracted DNA, the PCR primers we ordered, and a PCR program we made which followed typical protocol to run a PCR reaction. PCR requires multiple, ordered cycles of denaturation, annealing and extension, of which different temperatures and time periods are used. The denaturation step melts the DNA, causing the DNA to split into single strands. The annealing step then lowers the temperature enough for the primers we selected to bind to the now single stranded DNA at a specific location in the sequence. The extension step then allows polymerase to add dNTPs to the growing complementary strand, effectively adding complementary strands starting from our primers. This cycle must be run continually for many cycles to amplify our gene’s DNA. After the PCR reaction we ran a 1% Agarose gel electrophoresis to confirm that it was successful and that our DNA was amplified. This allows us to compare bands of PCR product DNA to molecular weight standards to identify our DNA and PCR primer in the gel. We then cut out the band we had identified as our amplified DNA and purified it. This gave us extremely pure DNA for our gene which we then sent to a lab to be sequenced.
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