Kun:LabNotes/ASE/2007-11-9: Difference between revisions

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(New page: ===Exp.1 eSNPs capture (cont.)=== Perform PCR on 5ul capturing reactions. x 1 x 12 template: 5ul each ABI 10x buffer: 4...)
 
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94C 3min -> 10 cycles of (94C 45sec -> 56C 2min -> 72C 1min) -> 20 cycles of (94C 45 sec -> 72C 1min) -> 72C 3min -> 15C hold.
94C 3min -> 10 cycles of (94C 45sec -> 56C 2min -> 72C 1min) -> 20 cycles of (94C 45 sec -> 72C 1min) -> 72C 3min -> 15C hold.
Repeat PCR on 10ul capturing reactions using Invitrogen's Platinum Taq.
                          x 1        x 12
      template:          10ul        each
      ABI 10x buffer:      9ul        108ul
      25mM MgCl2:          6ul        72ul
      10mM dNTP:          2ul        24ul
      100uM AmpF2Sol:    0.3ul        3.6ul
      100uM AmpRSol:    0.3ul        3.6ul
      50X SYBG I:        0.6ul        9.6ul
      iTaq:              0.5ul          6ul
      H2O:              72.4ul        869ul
94C 3min -> 10 cycles of (94C 45sec -> 56C 2min -> 72C 1min) -> 13 cycles of (94C 45 sec -> 72C 1min) -> take out the gDNA tube -> 9 cycles of (94C 45 sec -> 72C 1min)-> 72C 3min -> 15C hold.

Revision as of 22:56, 9 November 2007

Exp.1 eSNPs capture (cont.)

Perform PCR on 5ul capturing reactions.

                          x 1        x 12
     template:            5ul         each
     ABI 10x buffer:    4.5ul         54ul
     25mM MgCl2:          3ul         36ul
     10mM dNTP:           1ul         12ul
     100uM AmpF2Sol:   0.15ul        1.8ul
     100uM AmpRSol:    0.15ul        1.8ul
     50X SYBG I:        0.4ul        4.8ul
     iTaq:              0.5ul          6ul
     H2O:              35.3ul        430ul

94C 3min -> 10 cycles of (94C 45sec -> 56C 2min -> 72C 1min) -> 20 cycles of (94C 45 sec -> 72C 1min) -> 72C 3min -> 15C hold.

Repeat PCR on 10ul capturing reactions using Invitrogen's Platinum Taq.

                          x 1        x 12
     template:           10ul         each
     ABI 10x buffer:      9ul        108ul
     25mM MgCl2:          6ul         72ul
     10mM dNTP:           2ul         24ul
     100uM AmpF2Sol:    0.3ul        3.6ul
     100uM AmpRSol:     0.3ul        3.6ul
     50X SYBG I:        0.6ul        9.6ul
     iTaq:              0.5ul          6ul
     H2O:              72.4ul        869ul

94C 3min -> 10 cycles of (94C 45sec -> 56C 2min -> 72C 1min) -> 13 cycles of (94C 45 sec -> 72C 1min) -> take out the gDNA tube -> 9 cycles of (94C 45 sec -> 72C 1min)-> 72C 3min -> 15C hold.