Daniel:Notebook/ComboLock/2017-4-12
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RCA Double Deuce
Protocol
- Bglii digestion
- Make the following reactions; Do not add Bglii yet
- Aliquot 14 uL master mix into new tube
- Add 5 uL appropriate sample
- Use the following thermocycler program
- 2 hr 37C; When this cycle starts add 1 uL Bglii
- Heat kill with 65C for 20 min
Reagent | Single Rxn Vol | Master Mix (13.2X) |
RCA Rxn | 5 | 0 |
Bglii Cutter Primer + (100 uM) | 2 | 26.4 |
10X Buffer 3.1 | 2 | 26.4 |
Bglii | 1 | 0 |
nfH2O | 10 | 132 |
Total | 20 | 184.8 |