Jie:LabNotes/CpgSeq/2009-6-29
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ASE validation of fuzzy methylated genes[edit]
validation of primers[edit]
I use the Hybrid1-ES(1.9ug/ul): 63.4ng/ul to test the primers. Hybrid1-ES preparation refer to LabNotes on [1] PCR 2x iQ supermix 20 x 17 Primer F (25uM) 0.5 0.5 each Primer R (25uM) 0.5 0.5 each Hybrid1-ES(1.9ug/ul): 63.4ng/ul 0.1 1.7 dH2O 20 340 total 40ul The primers are 20090623_fuzzymethyl_ASE_valid. primers refer to [2]
I set the annealing temperature at 58C, 62C. 96C 3min-> 40x (95C 30sec, 58C or 62C 1min, 72C 1min)-> 72C 5min.
File:20090630 fuzzy methyl ASE valid 1.jpg20090630_fuzzy_methyl_ASE_valid_1
File:20090630 fuzzy methyl ASE valid 2.jpg20090630_fuzzy_methyl_ASE_valid_2
validation of samples are performed on 2009-07-02. refer to [3]