Daniel:Notebook/ComboLock/2017-3-30: Difference between revisions
Jump to navigation
Jump to search
>Djacobse (Created page with "=RCA Test (Started March 24)= Back to Calendar") |
>Djacobse |
||
Line 1: | Line 1: | ||
=RCA Test (Started [[Daniel:Notebook/ComboLock/2017-3-24|March 24]])= | =RCA Test (Started [[Daniel:Notebook/ComboLock/2017-3-24|March 24]])= | ||
<ol> | |||
<li>Qiaquick Column</li> | |||
<ol type="A"> | |||
<li>Add 100 uL PB (binding buffer) to sample</li> | |||
<li>Load sample onto column and spin for 1 minute at 14000 rpm; discard flow through</li> | |||
<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 2 minutes at 14000 rpm; discard flow through</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>Add ≥30 uL EB to column</li> | |||
<li>Let stand 1 minute</li> | |||
<li>Spin for 1 minute at 14000 rpm</li> | |||
</ol> | |||
<li>TBE Gel</li> | |||
<ol type="A"> | |||
<li>Mix 72 uL TBE and 24 uL 6X dye</li> | |||
<li>Aliquot 8 uL on to parafilm per sample/ladder</li> | |||
<li>Add 4 uL reaction or 2 uL ladder to appropriate aliquot</li> | |||
<li>Add 10 uL mix to gel lanes</li> | |||
<li>Run gel for 24 minutes at 230V</li> | |||
<li>Stain with 2 uL SYBR gold for 3 minutes</li> | |||
<li>Rinse and image in gel doc</li> | |||
</ol> | |||
</ol> | |||
[[Daniel:Notebook/ComboLock|Back to Calendar]] | [[Daniel:Notebook/ComboLock|Back to Calendar]] |
Revision as of 22:48, 30 March 2017
RCA Test (Started March 24)
- Qiaquick Column
- Add 100 uL 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
- 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 EB to column
- Let stand 1 minute
- Spin for 1 minute at 14000 rpm
- TBE Gel
- Mix 72 uL TBE and 24 uL 6X dye
- Aliquot 8 uL on to parafilm per sample/ladder
- Add 4 uL reaction or 2 uL ladder to appropriate aliquot
- Add 10 uL mix to gel lanes
- Run gel for 24 minutes at 230V
- Stain with 2 uL SYBR gold for 3 minutes
- Rinse and image in gel doc