Daniel:Notebook/PosSequencing/2017-5-20
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Single Stranded Probe Binding Test (Started Yesterday)
Protocol
- Single primer qPCR
- Make the following master mixes
- Add 37 uL or 38 uL (dual) to the appropriate lanes
- Add 2 uL samples and 1 (or 2) uL primer according to plate layout File:PlateLayout-20170520-SinglePrimer.png
- qPCR Cycles
- 95C 3 min
- 95C 3 sec
- 55C 30 sec
- 72C 20 sec
- plate read
- goto b x30
- 72C 2 min
- 16C hold
Reagent | uL in Single Rxn | Master Mix P2 (3.2X) | Master Mix BioU (3.2X) | Master Mix Dual (2.2X) |
Primer 2 (10 uM) | 1 | 3.2 | 3.2 | 2.2 |
BioU Primer (10 uM) | 1 | 0 | 0 | 2.2 |
2X Kapa SYBR Master Mix | 20 | 64 | 64 | 44 |
nfH2O | 17 | 54.4 | 54.4 | 35.2 |
Total | 121.6 | 121.6 | 83.6 |