Daniel:Notebook/ComboLock/2017-2-1: Difference between revisions
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=RCA Time Test= | =RCA Time Test (Started [[Daniel:Notebook/ComboLock/2017-1-27|Friday Jan 27]])= | ||
[[Daniel:Notebook/ComboLock|Back to Calendar]] | [[Daniel:Notebook/ComboLock|Back to Calendar]] | ||
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<li>Incubate at 65C for 10 minutes;Hold at 10C until next step</li></ol> | <li>Incubate at 65C for 10 minutes;Hold at 10C until next step</li></ol> | ||
Continued [[Daniel:Notebook/ComboLock/2017-2-2|tomorrow]] | |||
[[Category:ComboLock]] [[Category:20170127]] | |||
[[Category:ComboLock]] [[Category: |
Latest revision as of 17:17, 3 February 2017
RCA Time Test (Started Friday Jan 27)[edit]
Phosphorothiorate RCA Primer[edit]
- Rolling Circle Amplification
- Prepare master mix according to table below
- Add 14 uL master mix to each tube
- Add sample and to new reaction tubes according to following table
- Incubate at 30C for 3 hours
- Incubate at 65C for 10 minutes;Hold at 10C until next step
Reagent | uL Added | Master Mix (8.2X) |
Template | 3 | NA |
RCA Primer (10 uM) | 2.5 | 20.5 |
dNTP (1 mM) | 5 | 41 |
10X Buffer | 2 | 16.4 |
Phi29 | 1 | 0 |
BSA (10 mg/mL) | 0.4 | 3.28 |
nfH2O | 6.1 | 50.02 |
Total | 20 | 131.2 |
Sample Code | Phi29 Source | Incubation Time (hrs) | Sample (1AB) | No C Probes (3AB) |
N | NEB | 3 hrs | X | X |
O | NEB | 3 hrs-Epicentre | X | X |
Continued tomorrow