Daniel:Notebook/ComboLock/2017-3-30: Difference between revisions
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'''Gel Result''' | '''Gel Result''' | ||
[[Image:|600px]] | [[Image:2017-03-30-RCATest-CPLP-Bglii-TF-3hr-Purified.png|600px]] | ||
[[Daniel:Notebook/ComboLock|Back to Calendar]] | [[Daniel:Notebook/ComboLock|Back to Calendar]] |
Revision as of 23:17, 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
Gel Result