Daniel:Notebook/ComboLock/2017-4-12: Difference between revisions
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<li>Aliquot 12 uL master mix into new tube</li> | <li>Aliquot 12 uL master mix into new tube</li> | ||
<li>Add 7 uL appropriate sample</li> | <li>Add 7 uL appropriate sample</li> | ||
<li> | <li>Mix with vortexer, spin down, and add 1 uL Bglii</li> | ||
<li>Incubate 2 hr 37C</li> | |||
<li>2 hr 37C | |||
<li>Heat kill with 65C for 20 min</li></ol> | <li>Heat kill with 65C for 20 min</li></ol> | ||
</ol> | </ol> |
Revision as of 15:14, 12 April 2017
RCA Double Deuce
Protocol
- Bglii digestion
- Make the following reactions; Do not add Bglii yet
- Aliquot 12 uL master mix into new tube
- Add 7 uL appropriate sample
- Mix with vortexer, spin down, and add 1 uL Bglii
- Incubate 2 hr 37C
- Heat kill with 65C for 20 min
Reagent | Single Rxn Vol | Master Mix (13.2X) |
RCA Rxn | 7 | 0 |
10X Buffer 3.1 | 2 | 26.4 |
Bglii | 1 | 0 |
nfH2O | 10 | 132 |
Total | 20 | 158.4 |