Daniel:Notebook/ComboLock/2016-11-16: Difference between revisions
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>Djacobse (Created page with "=BSA Beads Test (Started Yesterday)= Back to Calendar ==Protocol== <li>Rolling Circle Amplification</...") |
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==Protocol== | ==Protocol== | ||
<ol start="5"> | |||
<li>Rolling Circle Amplification</li> | <li>Rolling Circle Amplification</li> | ||
<ol type="A"> | <ol type="A"> | ||
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<li>qPCR</li> | <li>qPCR</li> | ||
</ol> | </ol> | ||
=Library Prep (Started [[Daniel:Notebook/ComboLock/2016-11-15|Yesterday]]; Exp. Start [[Daniel:Notebook/ComboLock/2016-11-7|11/7/16]])= | |||
<ol start="2"> | |||
<li>Ethanol Precipitation</li> | |||
<ol type="A" start="2"> | |||
<li>Centrifuge for 25 minutes at 12000 rpm at 4C</li> | |||
<li>Remove supernatant and add 750 uL 70% EtOH</li> | |||
<li>Centrifuge for 12 minutes at 12000 rpm at 4C</li> | |||
<li>Remove supernatant and spin in vacuum centrifuge for 10 minutes</li> | |||
<li>Add 30 (20 for NTC) uL nfH2O to resuspend; pool samples A and B by reusing the water</li> | |||
</ol></ol> |
Revision as of 18:13, 16 November 2016
BSA Beads Test (Started Yesterday)
Protocol
- Rolling Circle Amplification
- Prepare 10.2X Master mix according to table below
- Add sample to new reaction tubes according to following table
- Incubate at 37C for 3 hours
- Incubate at 65C for 10 minutes;Hold at 10C until next step
- qPCR
Reagent | uL Added | Master Mix (10.2X) |
Template | 5 | NA |
RCA Primer (100 uM) | 2.5 | 25.5 |
dNTP (1 mM) | 0.8 | 8.16 |
10X Buffer | 2 | 20.4 |
Phi29 | 1 | 10.2 |
nfH2O | 8.7 | 88.74 |
Total | 20 | 153 |
Library Prep (Started Yesterday; Exp. Start 11/7/16)
- Ethanol Precipitation
- Centrifuge for 25 minutes at 12000 rpm at 4C
- Remove supernatant and add 750 uL 70% EtOH
- Centrifuge for 12 minutes at 12000 rpm at 4C
- Remove supernatant and spin in vacuum centrifuge for 10 minutes
- Add 30 (20 for NTC) uL nfH2O to resuspend; pool samples A and B by reusing the water