Daniel:Notebook/ComboLock/2017-5-2: Difference between revisions
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>Djacobse (Created page with "=Production Run (Started Friday 4/28)= Back to Calendar ==Library Prep== <ol start="10"> <li>Qiaquick ...") |
>Djacobse |
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<li>Add 200 uL (5X) PB (binding buffer) to sample</li> | <li>Add 200 uL (5X) PB (binding buffer) to sample</li> | ||
<li>Load sample onto column and spin for 1 minute at 14000 rpm; discard flow through</li> | <li>Load sample onto column and spin for 1 minute at 14000 rpm; discard flow through</li> | ||
<li>Add | <li>Add 700 uL PE (wash buffer) to column and spin for 1 minute at 14000 rpm; discard flow through</li> | ||
<li>Dry spin column for | <li>Repeat above step with 400 uL PE buffer</li> | ||
<li>Dry spin column for 2 minutes at 14000 rpm; discard flow through</li> | |||
<li>Let stand with cover open in fume hood for ~5 minutes</li> | <li>Let stand with cover open in fume hood for ~5 minutes</li> | ||
<li>Transfer column to a new 1.5 mL eppendorf tube</li> | <li>Transfer column to a new 1.5 mL eppendorf tube</li> | ||
<li>Add | <li>Add 30 uL nfH<sub>2</sub>O to column</li> | ||
< | |||
<li>Spin for 1 minute at 14000 rpm</li> | <li>Spin for 1 minute at 14000 rpm</li> | ||
</ol> | </ol> |
Revision as of 16:34, 2 May 2017
Production Run (Started Friday 4/28)
Library Prep
- Qiaquick Purification
- Combine 20 uL each sample into a 1.5 mL tube following matrix
- Add 200 uL (5X) PB (binding buffer) to sample
- Load sample onto column and spin for 1 minute at 14000 rpm; discard flow through
- Add 700 uL PE (wash buffer) to column and spin for 1 minute at 14000 rpm; discard flow through
- Repeat above step with 400 uL PE buffer
- Dry spin column for 2 minutes at 14000 rpm; discard flow through
- Let stand with cover open in fume hood for ~5 minutes
- Transfer column to a new 1.5 mL eppendorf tube
- Add 30 uL nfH2O to column
- Spin for 1 minute at 14000 rpm
- Size Select TBE Gel
Tube | Sample 1 | Sample 2 | Indexes |
1X | 1AX | 1BX | 20 |
5AX | 5AX | nfH2O | 24 |
1Y | 1AY | 1BY | 25 |
5AY,4AY | 5AY | 4AY | 26,27 |