Lab 10 – Erythranthe guttata PCR set up

In this week’s lab, we did a PCR run on our samples, using the following methods.

1. We partnered up and labeled 6 PCR tubes P1-P6 (for Pyrimidudes 1-6), noting which label corresponds to which samples. Ours were labeled:

P1 – HWH CATB

P2 – MAPL 002

P3 – SHOR 001

P4 – NPI-1

P5 – MONO 007

P6 – PRBW 006

1. We micro-pipetted 1 microliter of our samples of extracted DNA into the appropriate PCR tube.
2. We created a master mix enough for 16 PCR reactions, for our lab group to share. This master mix consisted of:
   1. ddH2O (214 microliters)
   2. 10x buffer (32 microliters)
   3. MgCl2 (32 microliters)
   4. BSA (16 microliters)
   5. dNTPs (3.2 microliters)
   6. Forward primer (3.2 microliters)
   7. Reverse primer (3.2 microliters)
   8. Taq (0.64 microliters)
3. We vortexed the master to mix it up. Then we added 19 microliters of the master mix to each of the PCR tubes with the DNA in them.
4. These PCR tubes were then added to the thermocycler to run PCR.