Daniel:Notebook/ComboLock/2017-1-25
Jump to navigation
Jump to search
Protocol
- Make qPCR master mix according to following recipe
- 361.2 uL nfH2O
- 17.2 uL AmpF-CLv3
- 17.2 uL AmpR-CLv3
- 430 uL Kapa SYBR Fast
- Add 48 uL master mix to each well
- Add 2 uL sample 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
- Mix 80 uL TBE, 20 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 22 minutes at 250V
- Open gel and stain with 3 uL SYBR Gold for 3 minutes
- Rinse gel and image in gel doc