Procedure Digital PCR
- 16 PCR Reactions were run today, following the procedure listed below.
- TNF A forward and reverse primers were diluted from 100uM to 10uM. Initial stock DNA (Concentration = 2.3mg/mL) was diluted several times to obtain 4 different templates, listed below:
- D0 = U20S 1:100 = 6.24ng/mL
- D1 = 0.624ng/mL
- D2 = 0.0624ng/mL = 6.24*10^(-2) ng/mL
- D3 = 6.24pg/mL
- For each PCR reaction, the following volumes will be added:
- 1uL of forward TNF A primer
- 1uL of reverse TNF A primer
- 1uL of diluted DNA (either U20S 1:100, D1, D2, or D3 for each sample)
- 50uL of PCR master mix
- 47 uL of dH20
- 8 samples of each template will be put through the PCR sequence (NOTE: The program to be run is named "First Trial 121713")
- 2 samples of plain water (instead of template) will also be tested as a negative control.
PCRs Run Today
- 8 D0 wells
- 8 D1 wells
- Labeled and stored in freezer.