Jie:LabNotes/ASE/2008-9-22

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Revision as of 17:42, 22 September 2008 by KunZhang (talk | contribs) (New page: ==padlock capture of sample from Yu Gao with eSNP84K== capture with the padlock probe made on 08-03-27. tube 1: G2 (~0.25ug/ul) tube 2: 2B (~3.5ng/ul) tube 3: 3B (~3.5ng/ul) tu...)
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padlock capture of sample from Yu Gao with eSNP84K

capture with the padlock probe made on 08-03-27.

 tube 1: G2 (~0.25ug/ul)
 tube 2: 2B (~3.5ng/ul)
 tube 3: 3B (~3.5ng/ul)
 tube 4: H1 (~5ng/ul)
 tube 5: H2 (~100ng/ul)
 tube 6: Jurkat DNA (5ng/ul)
 tube 7: negative control
 
 reaction system                 tube 1          x7 
 H2O                              6.3ul        44.1ul
 10x Ampligase Buffer               1ul           7ul
 probe(2008-03-27,30ng/ul)          3ul          21ul
 template                           2ul          2ul each
                                                                    ddH2O     Ligase Buffer   Ligase     Amplitaq    dNTP  
 2U/ul AmpliTaq Stoffel fragment; 0.5U/ul AmpLigase; 0.8nM dNTP      0.2ul         0.1ul       0.1ul       0.2ul    (2nM)0.4ul    
                                                              x20      4ul           2ul         2ul         4ul     8ul

Reaction program:

  95c 10min -> 63C 14h -> add 1ul SLN mix(2U/ul AmpliTaq Stoffel fragment; 0.5U/ul AmpLigase; 0.8nM dNTP) -> 60C 4h -> 
12 cycels (95C 1min -> 0.1C/sec to 60C, 60C for 4h 55C 1h) -> 95C 1min -> 37C 1min ->
add 2ul Exonuclease I/III mix -> 37C 2h -> 94C 5min -> 4C hold.