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...) |
>Jie deng No edit summary |
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===PCR on bisuphite conversed Jurkat-DNA=== | ===PCR on bisuphite conversed Jurkat-DNA=== | ||
2x iQ supermix 20 x | 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. | BS Jurkat gDNA (202ng/ul) 0.1 1.2 | ||
dH2O 20 | 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.