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

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==RE-amplify CViB sets for Covaris shearing==
==RE-amplify CViB sets for Covaris shearing==
*previously captured libraries on CViB sets
*previously captured libraries on CViB and PGP7 sets
*the protocol used for the previous steps can be found under labnotes 12-18-09 and 1-5-10
*the protocol used for the previous steps can be found under labnotes 12-18-09 and 1-5-10 (for CViB sets) and 9-8-09 and 10-26-09 (for PGP7)
  PCR
  PCR
  Template:          0.25ul
  Template:          0.25ul

Revision as of 22:02, 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 on CViB and PGP7 sets
  • the protocol used for the previous steps can be found under labnotes 12-18-09 and 1-5-10 (for CViB sets) and 9-8-09 and 10-26-09 (for PGP7)
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.