Daniel:Notebook/ComboLock/2017-4-7: Difference between revisions
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>Djacobse (Created page with "=RCA Test CPLP (Started March 24)= Back to Calendar ==Bgl Digestion-Minus Strand== <ol> <li>Bglii Dige...") |
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<li>Heat kill with 65C for 20 min</li></ol> | <li>Heat kill with 65C for 20 min</li></ol> | ||
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[[Image:2017-04-07-RCATest-CPLP-Bglii_PlusMinus.png|500px]] |
Latest revision as of 19:26, 7 April 2017
RCA Test CPLP (Started March 24)[edit]
Bgl Digestion-Minus Strand[edit]
- Bglii Digestion
- Make the following reactions
- Aliquot 15 uL master mix to appropriate tubes
- Aliquot 4 uL reaction to appropriate tubes: use QB, QC, QE, QF, XB, XC, XE, XF
- Use the following thermocycler program
- 5 min 95C
- Ramp to 50C at 0.2C/s
- 10 min 50C
- 1 hr 37C; When this cycle starts add 1 uL Bglii
- Heat kill with 65C for 20 min
Reagent | Single Rxn Vol | Master Mix + (4.2X) | Master Mix - (4.2X) |
RCA Rxn | 4 | 0 | 0 |
RCA Primer + (10 uM) | 2 | 8.4 | 0 |
RCA Primer + (10 uM) | 2 | 0 | 8.4 |
10X Buffer 3.1 | 2 | 8.4 | 8.4 |
Bglii | 1 | 0 | 0 |
nfH2O | 11 | 46.2 | 46.2 |
Total | 20 | 63 | 63 |