AlanFung:LabNotes/Sequencing/2009-11-24: Difference between revisions

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>Alan6017518
(New page: ==Sequencing Library Construction with Single Adaptor Protocol== ==PCR with dUTP and 6.3NH2 Primer== {| {{table}} | ||1 well |- | H2O||46.6 |- | Amp F6.3NH2(100uM)||0.2 |- | Amp R6.3NH2 (1...)
 
>Alan6017518
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==Sequencing Library Construction with Single Adaptor Protocol==
==Sequencing Library Construction with Single Adaptor Protocol-CVF-N Set==
 
==PCR with dUTP and 6.3NH2 Primer==
==PCR with dUTP and 6.3NH2 Primer==
{| {{table}}
{| {{table}}

Revision as of 18:56, 24 November 2009

Sequencing Library Construction with Single Adaptor Protocol-CVF-N Set

PCR with dUTP and 6.3NH2 Primer

1 well
H2O 46.6
Amp F6.3NH2(100uM) 0.2
Amp R6.3NH2 (100uM) 0.2
dUTP (1mM) 1.6
50X SYBG I 0.4
2X Master Mix 50
Template 1
Total 100
  • Qiaquick purification, elute with 32ul

USER Digestion

DNA 30
USER 3
Total 33

S1 Nuclease Digestion

S1 Digestion
10X S1 Nuclease Buffer 4
DNA after USER Digestion 33
S1 Nuclease (10U/ul) 1
ddH2O 2
  • 37C 10mins
  • Minelute purification. Elute w/16uH2O

End Repair