Kun:LabNotes/ASE/2008-1-8

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Exp.1 Capturing cSNPs with the eSNP84k probe set(repeat)[edit]

Templates:

  1. NA20431, 347ng/ul, one reaction with the equal molar pool of all five probe sets, 800ng/reaction.
  2. GM20431 ds-cDNA, 122ng/ul, Lib #1: 1ng; Lib #2: 20ng; Lib #3 50ng; Lib #4: 100ng; Lib #5: 400ng
  3. GC1EP, 114ng/ul, Lib #1: 10ng; Lib #2: 20ng; Lib #3 50ng; Lib #4: 100ng; Lib #5: 400ng
NA20431 GM20431 GC1EP
Template 2.3 1(20ng) 1(20ng) 1(50ng) 0.85 3.4 1(20ng) 1(20ng) 1(50ng) 0.88 3.5
10x buffer 1.5 1 1 1 1 1 1 1 1 1 1
Lib #1 (384nM) 2 2 2
Lib #2 (580nM) 1.5 1.5 1.5
Lib #3 (331nM) 2.6 2.6 2.6
Lib #4 (500nM) 2.4 2.4 2.4
Lib #5 (460nM) 1.8 1.8 1.8
H2O 0.9 6 6.5 5.4 5.75 3.8 6 6.5 5.4 5.72 3.7

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.