Daniel:Notebook/ComboLock/2017-3-30: Difference between revisions
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=RCA Test (Started [[Daniel:Notebook/ComboLock/2017-3-24|March 24]])= | =RCA Test (Started [[Daniel:Notebook/ComboLock/2017-3-24|March 24]])= | ||
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Revision as of 19:39, 3 April 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 File:20170330-GelLanes-Purified.png
- Run gel for 24 minutes at 230V
- Stain with 2 uL SYBR gold for 3 minutes
- Rinse and image in gel doc
Gel Result