Jie:LabNotes/CpgSeq/2009-6-29

From ZhangLabWiki
Jump to navigation Jump to search

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]