AlanFung:Protocol/Construction of Solexa sequencing Library(USER)

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Sequencing Library Construction with Single Adaptor Protocol[edit]

PCR with dUTP and 6.3NH2 Primer[edit]

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[edit]

DNA 30
USER 3
Total 33

S1 Nuclease Digestion[edit]

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[edit]