For the Pyrimidudes, Lab 4 was all about separating out and then cleaning up the products of our PCR reactions from the last lab section. In order to set up the gel, the Pyrimidudes planned out which samples would go where; in total, we had 13 samples, plus a ladder and control. In order to prep out PCR, products, I pipetted out 15 1.5 microliter dots of loading dye, while the rest of the Pyrimidudes carefully set up the PCR reactions for adding to the dots. Each of our group added 3 microliters of DNA to each dye dot, with the exception of myself, who added the dye dot and PCR products directly into each lane, where they were mixed in-situ by pipetting up and down. We then run the gel at 130 volts for 30 minutes.
As this was running, we next prepped at PCR cleanup using ExoSAP of some of the remaining samples by creating a master mix using 190.62 μL H2O, 22.5 μL 10x SAP Buffer, 7.9 μL EXP, and 3.96 μL SAP for a total volume of 225 μL. We then added 12.5 μL of this master mix to each of 13 different labeled PCR tubes, to which was added 7.5 μL each of our PCR products. The PCR products were then run for 45 minutes by Dr. Paul using the ExoSAP protocol.