3/27 PCR Day/Making Gels

Today we prepared reactions and ran PCR. We are testing two different types of DNA (Oats and GMO Positive DNA, and are using two different primers (one that tests for PSII and one that tests for GMOs).This results in a total of four reactions. We ran 25 uL reactions, each consisting of 2.5 uL 10x Buffer, .5 uL dNTPs, 1 uL Primers, 10 uL DNA, .125 uL Taq, and 11 uL H2O. We loaded all of the reactions into the PCR machine and ran the reactions as follows:

94 degrees for 2 mins

40 cycles of:

94 degrees for 1 min

59 degrees for 1 min

72 degrees for  2 min

72 degrees for 10 mins

Hold at 15 degrees.


PCR of Samples

We also poured a 0.7% Agarose Gel. We used a hot-plate to melt the already prepared 0.7% agarose mixture that we retrieved from the fridge and placed the Erlenmeyer flask with the now-liquid agarose in a beaker of cold water to bring down the temperature of the liquid. Then, we poured the liquid into the gel tray with one comb and placed the gel tray in the fridge to set.


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