Kun:LabNotes/ASE/2008-1-12
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Exp.1 Capturing cSNPs with the eSNP84k probe set(repeat)[edit]
Templates:
- Hues6 120ng/ul, one reaction with the equal molar pool of all five probe sets, 800ng/reaction.
- Hues6 ds-cDNA, 53ng/ul, Lib #1: 10ng; Lib #2: 20ng; Lib #3 50ng; Lib #4: 100ng; Lib #5: 200ng
- GC1F, 167ng/ul, Lib #1: 10ng; Lib #2: 20ng; Lib #3 50ng; Lib #4: 100ng; Lib #5: 400ng
Hues6 | Hues6 | GC1F | |||||||||
Template | 6.5 | 1(20ng) | 1(20ng) | 1(50ng) | 0.85 | 3.4 | 1(20ng) | 1(20ng) | 1(50ng) | 0.88 | 3.5 |
10x buffer | 2.5 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
Lib #1 (384nM) | 3.4(223nM) | 3.4(223nM) | 2 | ||||||||
Lib #2 (580nM) | 3(281nM) | 3(281nM) | 1.5 | ||||||||
Lib #3 (331nM) | 2.6 | 2.6 | 2.6 | ||||||||
Lib #4 (500nM) | 3.9(308nM) | 3.9(308nM) | 2.4 | ||||||||
Lib #5 (460nM) | 3.4(246nM) | 3.4(246nM) | 1.8 | ||||||||
H2O | 0 | 4.6 | 5 | 5.4 | 3.1 | 1.6 | 6 | 6.5 | 5.4 | 5.9 | 4.8 |
94C 3min->60C 40h-> add 1.5ul/1.0ul SLN mix (1U/ul stoffel, 0.5U/ul AmpLigase, 200nM dNTP) -> 60C 1h -> 15 cycles of (94C 1min -> 60C 1h) -> 60C hold -> 37C 1min -> add 2ul of exo mix (Exo I, Exo III, T7 exo)->37c 2 hour -> 94C 5min -> 4C hold.