Kun:LabNotes/CpgSeq/2008-3-18: Difference between revisions
Jump to navigation
Jump to search
(New page: ==Exp. 1 Cpg capture with a new batch of bisulfite treated Jurkat DNA from 03/17/08== Sigma x2 Zymo x 2 Template 7 3 10X buffer(fre...) |
No edit summary |
||
Line 13: | Line 13: | ||
H2O 0 | H2O 0 | ||
94C 10min -> 60C 10min -> add 1ul NSL mix (Stoffel 2U/ul, Ampligase 0.5U/ul, dNTP 10nM or 50uM) -> 60C 1h -> | 94C 10min -> 60C 10min -> add 1ul NSL mix (Stoffel 2U/ul, Ampligase 0.5U/ul, dNTP 10nM or 50uM) -> 60C 1h -> 45 cycles of (94C 1min -> 60C 1h) -> 60C hold -> 37C 1min -> add 2ul Exo I&III, 37C 2h -> 94C 5min -> 4C hold. | ||
===PCR=== | |||
Template 12ul | |||
2X iProof Mastermix 100ul | |||
100uM AmpF6.2 0.8ul | |||
100uM AmpR6.2 0.8ul | |||
50X SYBG I 0.8ul | |||
H2O 85ul | |||
Split into two tubes. | |||
98C 30S -> (98C 10S -> 58C 20S -> 72C 20S) x 15 | |||
-> 72C 5 min -> 15C hold. |
Revision as of 17:09, 21 March 2008
Exp. 1 Cpg capture with a new batch of bisulfite treated Jurkat DNA from 03/17/08
Sigma x2 Zymo x 2 Template 7 3 10X buffer(fresh) 1 1 Cpg28k probe(218nM) 2 2 H2O 0 4
Exon capture control
GM20431 131ng/ul 7 10X buffer(fresh) 1 E55kv1 probe(220nM) 2 H2O 0
94C 10min -> 60C 10min -> add 1ul NSL mix (Stoffel 2U/ul, Ampligase 0.5U/ul, dNTP 10nM or 50uM) -> 60C 1h -> 45 cycles of (94C 1min -> 60C 1h) -> 60C hold -> 37C 1min -> add 2ul Exo I&III, 37C 2h -> 94C 5min -> 4C hold.
PCR
Template 12ul 2X iProof Mastermix 100ul 100uM AmpF6.2 0.8ul 100uM AmpR6.2 0.8ul 50X SYBG I 0.8ul H2O 85ul Split into two tubes. 98C 30S -> (98C 10S -> 58C 20S -> 72C 20S) x 15 -> 72C 5 min -> 15C hold.