Kun:LabNotes/CpgSeq/2008-5-14: Difference between revisions

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   dNTP: 4uM
   dNTP: 4uM


   Reactions                            A            B (KlenTaq)            C
   Reactions                            A            B (KlenTaq)            C                     D
   H2O                                4.5ul        4.5ul                    0ul
   H2O                                4.5ul        4.5ul                    0ul                 4.5ul
   10X AmpLigase buffer                1ul          1ul                    1ul
   10X AmpLigase buffer                1ul          1ul                    1ul                  1ul
   Probe (04/30/08, 192nM)              3ul          3ul  05/06/08(58nM)  7.5ul
   Probe (04/30/08, 192nM)              3ul          3ul  05/06/08(58nM)  7.5ul 03/10/08(218nM)  3ul
   Bis-converted HD50AA (157ng/ul)    1.5ul        1.5ul                  1.5ul
   Bis-converted HD50AA (157ng/ul)    1.5ul        1.5ul                  1.5ul                1.5ul
   SLN mix                              1ul          1ul                    1ul
   SLN mix                              1ul          1ul                    1ul                  1ul
    
    
   5x (94C2min -> cool down at 0.1C/sec to 55c -> 55C 4h)-> 94C 2min  
   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
     -> 37C 1min -> add 2ul exo I&II -> 37C 2h -> 94C 5min -> 4C hold

Revision as of 00:48, 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                     D
 H2O                                4.5ul        4.5ul                     0ul                 4.5ul
 10X AmpLigase buffer                 1ul          1ul                     1ul                   1ul
 Probe (04/30/08, 192nM)              3ul          3ul   05/06/08(58nM)  7.5ul  03/10/08(218nM)  3ul
 Bis-converted HD50AA (157ng/ul)    1.5ul        1.5ul                   1.5ul                 1.5ul
 SLN mix                              1ul          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