AlanFung:LabNotes/Sequencing/2009-11-19

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Quantification of New amplified Phi-X and all libraries

  • Dilute all samples by 10 fold
Samples
PhiX-Stock CVF PGP4 PGP10 PGP8C
PhiX-Stock CVF PGP4 PGP10 PGP8C
PhiX-Stock 1-7 PGP1 PGP6 NA12878 PGP8D
PhiX-Stock 1-7 PGP1 PGP6 NA12878 PGP8D
PhiX 8 PGP1 PGP8 PGP8A PGP8E
PhiX 8 PGP1 PGP8 PGP8A PGP8E
CVI 9 PGP1 PGP9 PGP8B N12
CVI 9 PGP1 PGP9 PGP8B N12
  • split every 50ul rxn into 2 wells
98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min
X20 X1.1
PhiX Control DNA (10nM) 1.000
Syb_FP5 (100uM) 0.200 4.000 4.400
Syb_RP7 (100uM) 0.200 4.000 4.400
2X Phusion 25.000 500.000 550.000
50X SYBR Green I 0.200 4.000 4.400
H20 24.000 480.000 528.000