Daniel:Notebook/ComboLock/2017-2-1: Difference between revisions
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>Djacobse (Created page with "=RCA Time Test= Back to Calendar ==Phosphorothiorate RCA Primer== <ol> <li>Rolling Circle Amplification</li> <ol type="A"> <li>Prepare master ...") |
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<li>Add 14 uL master mix to each tube</li> | <li>Add 14 uL master mix to each tube</li> | ||
<li>Add sample and to new reaction tubes according to following table</li> | <li>Add sample and to new reaction tubes according to following table</li> | ||
<li>Incubate at 30C for | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="background-color:#B1A0C7;font-size:12pt;font-weight:bold" align="center" | |||
| width="85" height="45" | Sample Code | |||
| width="85" | Phi29 Source | |||
| width="98" | Incubation Time (hrs) | |||
| width="65" | Sample (1AB) | |||
| width="65" | No C Probes (3AB) | |||
|- style="font-size:12pt" align="center" | |||
| height="15" | N | |||
| NEB | |||
| 3 hrs | |||
|style="font-weight:bold" | X | |||
|style="font-weight:bold" | X | |||
|- style="background-color:#BFBFBF;font-size:12pt" align="center" | |||
| height="15" | O | |||
| NEB | |||
| 3 hrs-Epicentre | |||
|style="font-weight:bold" | X | |||
|style="font-weight:bold" | X | |||
|} | |||
<li>Incubate at 30C for 3 hours</li> | |||
<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> | ||
Revision as of 00:47, 2 February 2017
RCA Time Test
Phosphorothiorate RCA Primer
- 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 |