AlanFung:LabNotes/Sequencing/2009-12-5

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qPCR Quantification

PGP4 PGP6 PGP8 PGP9 PGP8D PGP8E Dpn-CV-iF2 Dpn-CV-F2

Setup
Amp Phi-X (1nM) PGP6 PGP9 PGP8E Dpn-CV-iF2 (1/10)
Amp Phi-X (1nM) PGP6 PGP9 PGP8E Dpn-CV-iF2 (1/10)
Amp Phi-X (1nM) PGP6 PGP9 PGP8E Dpn-CV-iF2 (1/10)
Amp Phi-X (1nM) PGP6 PGP9 PGP8E Dpn-CV-iF2 (1/10)
PGP4 PGP8 PGP8D Dpn-CV-F2 (1/10)
PGP4 PGP8 PGP8D Dpn-CV-F2 (1/10)
PGP4 PGP8 PGP8D Dpn-CV-F2 (1/10)
PGP4 PGP8 PGP8D Dpn-CV-F2 (1/10)
x18 *1.1
DNA 2
Syb_FP5 (100uM) 0.2 3.6 3.96
Syb_RP7 (100uM) 0.2 3.6 3.96
2X Phusion 25 450 495
50X SYBR Green I 0.2 3.6 3.96
H20 24 432 475.2
Total 982.08
  • Dilute sample, 1ul in 9ul ddh2o
  • mix 2ul 1/10 sample with 98ul mix
  • aliquot 24ul to each well
98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min