Kun:LabNotes/CpgSeq/2008-3-18: Difference between revisions

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(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...)
 
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   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 -> 59 cycles of (94C 1min -> 60C 1h) -> 60C hold -> 37C 1min -> add 2ul Exo I&III, 37C 2h -> 94C 5min -> 4C hold.
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.