Daniel:Notebook/ComboLock/2017-1-26: Difference between revisions
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File:|qPCR curve | File:20170126-qPCR-RCATest-Epicentre.png|qPCR curve | ||
File:2017-01-26-RCATest-Epicentre.png|Gel image | File:2017-01-26-RCATest-Epicentre.png|Gel image | ||
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Revision as of 21:03, 26 January 2017
RCA Test (Started Monday)
Epicentre Test (Started Yesterday)
Sample Matrix
- qPCR
- Make 5.2X qPCR master mix according to following recipe
- 109.2 uL nfH2o
- 5.2 uL AmpF</li
- 130 uL 2X Kapa SYBR Fast Master Mix
- Add 47 uL master mix to each well
- Add 2 uL sample and 1 uL appropriate Reverse Primer (1 uM) according to table
- qPCR Cycles
- 95C 3 min
- 95C 3 sec
- 55C 30 sec
- 72C 20 sec
- plate read
- goto b x24
- 72C 2 min
- 16C hold
- TBE Gel
- Mix 64 uL TBE, 16 uL 6x loading dye
- Aliquot 10 uL per sample/ladder lane onto parafilm
- Add 2 uL of sample or ladder to correct drop
- Load 10 uL in to well
- Run gel for 23 minutes at 230V
- Open gel and stain with 3 uL SYBR Gold for 3 minutes
- Rinse gel and image in gel doc
- 20170126-qPCR-RCATest-Epicentre.png
qPCR curve
- 2017-01-26-RCATest-Epicentre.png
Gel image