Kun:LabNotes/CpgSeq/2008-5-14: Difference between revisions
Jump to navigation
Jump to search
m (New page: ==cpgMIP28k capture== *The primary goal of this experiment is to test whether the probe I made by DpnII/TspRI digestion on the Bst-hRCA amplicon works or not. The probe made on 04/29/08 Ex...) |
|||
Line 14: | Line 14: | ||
SLN mix 1ul 1ul 1ul | SLN mix 1ul 1ul 1ul | ||
5x (94C2min -> cool down at 0.1C/sec to 55c -> 55C 4h)-> 94C 2min -> 37C 1min -> add 2ul exo I&II -> 37C 2h -> 94C 5min -> 4C hold | 5x (94C2min -> cool down at 0.1C/sec to 55c -> 55C 4h)-> 94C 2min | ||
-> 37C 1min -> add 2ul exo I&II -> 37C 2h -> 94C 5min -> 4C hold |
Revision as of 00:36, 15 May 2008
cpgMIP28k capture
- The primary goal of this experiment is to test whether the probe I made by DpnII/TspRI digestion on the Bst-hRCA amplicon works or not. The probe made on 04/29/08 Exp2 (192nM) with the PCR-USER protocol is used as the positive control.
- The secondary goal is to compare Stoffel fragment with KlenTaq fragment (ClontTech Titanium Taq).
SLN mix: AmpLigase: 0.5U/ul Stoffel or Klentaq: 2U/ul dNTP: 4uM
Reactions A B (KlenTaq) C H2O 4.5ul 4.5ul 0ul 10X AmpLigase buffer 1ul 1ul 1ul Probe (04/30/08, 192nM) 3ul 3ul 05/06/08(58nM) 7.5ul Bis-converted HD50AA (157ng/ul) 1.5ul 1.5ul 1.5ul SLN mix 1ul 1ul 1ul 5x (94C2min -> cool down at 0.1C/sec to 55c -> 55C 4h)-> 94C 2min -> 37C 1min -> add 2ul exo I&II -> 37C 2h -> 94C 5min -> 4C hold