Alice:LabNotes/2010-1-21
Jump to navigation
Jump to search
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 sets
- the protocol used for the previous steps can be found under labnotes 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.