Daniel:Notebook/ComboLock/2017-4-3
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RCA Test (Started March 24)
I'll try qPCR on the RCA product again, this time on the NEB 12 hr samples (since I am running low on X samples), but for few cycles and with only the forward primer (which should be RC to the RCA product. This will be fed into Bglii digestion
- qPCR-Second strand synthesis
- Make the following 13.2X master mix
- 13.2 uL AmpF-6.4Sol (10 uM)
- 264 uL Kapa SYBR 2X Master Mix
- 224.4 uL nfH2O
- Aliquot 38 uL master mix into each lane
- Add 2 uL sample according to plate layout File:PlateLayout-20170403-RCATest-CPLP-ssSynthesis.png
- Run the following protocol
- 95C for 2 min
- 95C for 5 sec
- 55C for 30 sec
- 72C for 45 sec
- Goto (b) x20
- 72C for 2 min
- Qiaquick Column
- TBE Gel
- Bgl Digestion