Daniel:Notebook/PosSequencing/2017-6-2
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Emulsion PCR Test 3 (Started Wednesday)
Protocol-Part 2
- Single Primer qPCR
- Make the following master mixes
- Aliquot 38 uL master mix to appropriate lanes
- Add 2 uL sample according to plate layout below File:PlateLayout-20170601-SPqPCR-emPCRTest3-Round2.png
- Run the following thermocycler program
- 95C 3 min
- 95C 3 sec
- 55C 30 sec
- 72C 20 sec
- plate read
- goto b x150
- 72C 2 min
- 16C hold
Reagent | uL in Single Rxn | Master Mix P2 (5.2X) | Master Mix BioU (5.2X) | Master Mix Dual (5.2X) |
Primer 2 (10 uM) | 1 | 5.2 | 0 | 5.2 |
BioU Primer (10 uM) | 1 | 0 | 5.2 | 5.2 |
2X Kapa SYBR Master Mix | 20 | 104 | 104 | 104 |
nfH2O | 17 | 88.4 | 88.4 | 83.2 |
Total* (+ 2 uL sample=40) | 38 | 197.6 | 197.6 | 197.6 |