Daniel:Notebook/ComboLock/2017-2-3: Difference between revisions
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>Djacobse (Created page with "=Version 2 Oligos Round 2 (Started yesterday)= Back to Calendar ==Protocol-Part 4==") |
>Djacobse |
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[[Daniel:Notebook/ComboLock|Back to Calendar]] | [[Daniel:Notebook/ComboLock|Back to Calendar]] | ||
==Protocol-Part 4== | ==Protocol-Part 4-RCA== | ||
<ol start="8"> | |||
<li>Exonuclease Digestion</li> | |||
<ol type="A"> | |||
<li>Mix 15 uL Exo I (20U/uL) and 3 uL ExoIII (100U/uL)</li> | |||
<li>Add 2 uL to each sample, mix by swirling pipette tip</li> | |||
<li>Incubate for 1 hour at 37C</li> | |||
<li>Heat kill by incubating for 5 min at 95C</li> | |||
</ol> | |||
<li>Rolling Circle Amplification</li> | |||
<ol type="A"> | |||
<li>Prepare 7X Master mix according to table below</li> | |||
<li>Add 15 uL master mix to each tube</li> | |||
<li>Add sample to new reaction tubes</li> | |||
<li>Incubate at 37C for 3 hours</li> | |||
<li>Incubate at 65C for 10 minutes;Hold at 10C until next step</li></ol> |
Revision as of 17:14, 3 February 2017
Version 2 Oligos Round 2 (Started yesterday)
Protocol-Part 4-RCA
- Exonuclease Digestion
- Mix 15 uL Exo I (20U/uL) and 3 uL ExoIII (100U/uL)
- Add 2 uL to each sample, mix by swirling pipette tip
- Incubate for 1 hour at 37C
- Heat kill by incubating for 5 min at 95C
- Rolling Circle Amplification
- Prepare 7X Master mix according to table below
- Add 15 uL master mix to each tube
- Add sample to new reaction tubes
- Incubate at 37C for 3 hours
- Incubate at 65C for 10 minutes;Hold at 10C until next step