Daniel:Notebook/ComboLock/2016-8-29
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Positive Control Dilution Test (Started Friday)
Protocol
Today I'm testing the samples using the P2 and P12RC amplification primers. Yesterday included the original amplicon materials; this qPCR should be only the DNA still attached to beads.
- qPCR X2-Primers
- Set up 14 reaction with 2 uL sample, 25 uL KAPA, 22 uL nfH2O, 0.5 uL primer2 and 0.5 uL primer12RC
- Master Mix
- 352.5 uL KAPA SyberFast MM
- 310.2 uL nfH2O
- 7.05 uL primer2
- 7.05 uL primer12RC
- Add 48 uL Master Mix to each well
- qPCR Cycles
- 95C 3 min
- 95C 3 sec
- 55C 30 sec
- 72C 20 sec
- plate read
- goto b x32
- 72C 2 min
- 16C hold