Kun:LabNotes/ASE/2008-1-1

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Exp.1 Check the purity of cDNAs by QPCR[edit]

I use two sets of three PCR primer pairs (C1,F1,G1) in the PGP1VER2 plates, one set targeting cDNA and the other targeting gDNA.

  Samples to test:
  GM20431(12/31/07) 122ng/ul
  GC1EP (12/31/07)  114ng/ul
  Hues6 (12/31/07)   53ng/ul
  H1 (12/31/07)     123ng/ul
  H2 (12/31/07)      72ng/ul
  BJ (12/31/07)      62ng/ul
  Hues29(08/07/07)   31ng/ul
  Hues30(08/07/07)   35ng/ul
  H1 (11/??/07)      48ng/ul
  H2 (11/??/07)      35ng/ul
                                  x60
  Template(5ng/ul)      1ul      6x10
  2X Taq Master mix:   10ul       600
  2uM primer mix:       2ul        each
  50X SYBG I:         0.1ul        6
  H2O                 6.9ul       420

94C 2min -> (94C 30sec -> 57C 30sec -> 72C 45sec)x 40 -> 72C 3min -> 15C hold

  File:2008-01-01-gDNA-contamination-check.png

The gDNA clean-up protocol removed most of the DNA contamination. The H1/H2 hybrid RNA samples seem to be quite clean even without this protocol. Hues29 and Hues30 are contaminated.