Jie:LabNotes/CpgSeq/2009-6-22: Difference between revisions

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>Jie deng
(New page: ==Methyl_PCR for ASE validation== ===PCR on bisuphite conversed Jurkat-DNA=== 2x iQ supermix 20 x 15 Primer F (50uM) 0.5 0.5 ea...)
 
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===PCR on bisuphite conversed Jurkat-DNA===
===PCR on bisuphite conversed Jurkat-DNA===


  2x iQ supermix                        20            x  15          
  2x iQ supermix                        20            x  12          
  Primer F (50uM) 0.5         0.5 each
  Primer F (50uM) 0.5         0.5 each
  Primer R (50uM) 0.5            0.5 each
  Primer R (50uM) 0.5            0.5 each
  BS Jurkat gDNA (202ng/ul)            0.1   1.5
  BS Jurkat gDNA (202ng/ul)            0.1   1.2
  dH2O         20 300
  dH2O         20 240
  total         40ul
  total         40ul


  The primers are plate2_A1-H1. I set the annealing temperature at 55C, 58C, 62C.  
  The primers are plate2_A1-H1. I set the annealing temperature at 55C, 58C, 62C.  
  For iQ supermix: 96C 3min-> 40x (95C 30sec, 62C 1min, 72C 1min)-> 72C 5min.
  For iQ supermix: 96C 3min-> 40x (95C 30sec, 62C 1min, 72C 1min)-> 72C 5min.

Revision as of 17:34, 22 June 2009

Methyl_PCR for ASE validation

PCR on bisuphite conversed Jurkat-DNA

2x iQ supermix                         20             x   12          
Primer F (50uM)			0.5	        0.5 each
Primer R (50uM)			0.5             0.5 each
BS Jurkat gDNA (202ng/ul)     	        0.1	  	1.2
dH2O				         20		240
total				         40ul
The primers are plate2_A1-H1. I set the annealing temperature at 55C, 58C, 62C. 
For iQ supermix: 96C 3min-> 40x (95C 30sec, 62C 1min, 72C 1min)-> 72C 5min.