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qPCR

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qPCR is a test that shows gene expression over time. In Dukes lab, we run this test on a 384 well plate. Each well will end up with 2ul of cDNA or coding DNA and 8ul of master mix. This is obtained from a process called RT or reverse transcription. In qPCR, you must first dilute your cDNA with water. Then a master mix must be created for each thing you want to look at in qPCR. This master mix is made of water, a forward and reverse primer and a reagent. There is a guide to a mix solution for one individual sample so it must be multiplied by however many samples you are testing plus one or two for pipet error. For example, I was running 12 samples so I multiplied by 15 in order to be sure I had enough of the mixture. Every sample is mixed in their individual well, a film is placed over the plate and it is placed in the centrifuge. Finally it is put in the qPCR machine which analyzes the gene expression.

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