First PCR test
PCR samples from 10/12
Materials - uL per sample:
- 10 mM primer F - 1 uL
- 10 mM primer R - 1 uL
- BSA - 0.5 ul
- 2x biomix - 10 uL
- water - 5.5 uL
Make master mix (MM), 18 uL MM per tube, 2 uL DNA per tube. C+ = positive control (PM7) C- = negatie control (water)
Thermalcycler protocol -> LOBMINI volume = 50 uL
| Temp | Time |
|---|---|
| Denaturation | |
| 94 ºC | 3:00 |
| Annealing | x34 |
| 94 ºC | 0:30 |
| 46 ºC | 0:30 |
| 72 ºC | 1:00 |
| Extension | |
| 72 ºC | 5:00 |
| 15 ºC | inf |
Procedure:
- Add 18 uL MM
- 2 uL DNA to each tube
- Do not mix after adding DNA but vortex MM
Written on October 19, 2020