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.