Alice:LabNotes/2010-1-21: Difference between revisions

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  MmeI(2U/ul)                2ul
  MmeI(2U/ul)                2ul
  37C 1hr, 80C 20min
  37C 1hr, 80C 20min
==RE-amplify CViB sets for Covaris shearing==
*previously captured libraries from 12-18-09 and 1-5-10
PCR
Template:                            0.25ul
2x Phusion master mix:    50ul   
100uM AmpF6.3NH2        0.2ul   
100uM AmpR6.3NH2        0.2ul   
50X SYBG I:            0.4ul   
H2O                      50ul   
98C 30sec -> (98C 10sec -> 58C 20sec -> 72C 20sec) x 6 -> 72C 3min -> 4C hold.

Revision as of 22:00, 21 January 2010

MmeI digestion on CViB sequencing libraries

  • Previous library construction protocol can be found under labnotes 1-9-10
  • size selection protocol can found under labnotes 1-15-10
setup:
DNA(15nM)                   10ul
10x buffer 4                2ul
1mM SAM                     2ul
MmeI(2U/ul)                 2ul
37C 1hr, 80C 20min

RE-amplify CViB sets for Covaris shearing

  • previously captured libraries from 12-18-09 and 1-5-10
PCR
Template:                             0.25ul
2x Phusion master mix:    50ul    
100uM AmpF6.3NH2         0.2ul    
100uM AmpR6.3NH2        0.2ul    
50X SYBG I:             0.4ul    
H2O                      50ul    
98C 30sec -> (98C 10sec -> 58C 20sec -> 72C 20sec) x 6 -> 72C 3min -> 4C hold.