Jie:LabNotes/ASE/2008-9-22
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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.