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]
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]