In this week’s lab, we did a PCR run on our samples, using the following methods.
- 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
- We micro-pipetted 1 microliter of our samples of extracted DNA into the appropriate PCR tube.
- We created a master mix enough for 16 PCR reactions, for our lab group to share. This master mix consisted of:
- ddH2O (214 microliters)
- 10x buffer (32 microliters)
- MgCl2 (32 microliters)
- BSA (16 microliters)
- dNTPs (3.2 microliters)
- Forward primer (3.2 microliters)
- Reverse primer (3.2 microliters)
- Taq (0.64 microliters)
- 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.
- These PCR tubes were then added to the thermocycler to run PCR.