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Quantify libraries with Phi-X ladder qPCR

AL-Exome-CViB-Jan14-AB w/ 1/10 dilution, AL-Exome-CViB-Jan14-CDE w/ 1/10 dilution, Ida's library

Setup
1nm PhiX AL-Exome-CViB-Jan14-AB AL-Exome-CViB-Jan14-CDE
1nm PhiX AL-Exome-CViB-Jan14-AB AL-Exome-CViB-Jan14-CDE
1nm PhiX AL-Exome-CViB-Jan14-AB AL-Exome-CViB-Jan14-CDE
1nm PhiX AL-Exome-CViB-Jan14-AB AL-Exome-CViB-Jan14-CDE
Ida 1/10 AL-Exome-CViB-Jan14-AB 1/10 AL-Exome-CViB-Jan14-CDE
Ida 1/10 AL-Exome-CViB-Jan14-AB 1/10 AL-Exome-CViB-Jan14-CDE
Ida 1/10 AL-Exome-CViB-Jan14-AB 1/10 AL-Exome-CViB-Jan14-CDE
Ida 1/10 AL-Exome-CViB-Jan14-AB 1/10 AL-Exome-CViB-Jan14-CDE
2 Wells X13
DNA 2 26
Syb_FP5 (100uM) 0.2 2.6
Syb_RP7 (100uM) 0.2 2.6
2X Phusion 25 ,
50X SYBR Green 0.2 2.6
H2O 24 312
Total 345.8
   * Dilute sample, 1ul in 9ul ddh2o
   * mix 2ul 1/10 sample with 98ul mix in non-stick tube
   * aliquot 24ul to each well, for each sample use the same tip for every well (Pipet slowly!)

98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min

Media:JD-cpg97k+30k-Breast_cancer-01230B09-Jan15_log.pdf
Media:AL-Exome-CViB-Jan14-AB.pdf
Media:AL-Exome-CViB-Jan14-CDE.pdf

JD-cpg97k+30k-Breast_cancer-01230B09-Jan15

Well / Set C(t) Difference nM
PhiX A1 6.98 7.06
PhiX B1 7.2
PhiX C1 7.16
PhiX D1 6.89
JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 E1 6.82 6.92 0.14 1.1
JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 F1 7.09
JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 G1 6.89
JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 H1 6.86

AL-Exome-CViB-Jan14-AB

Well / Set C(t) Ave. Difference nM
PhiX A1 7.99 8.11
PhiX B1 8.33
PhiX C1 8.18
PhiX D1 7.95
AL-Exome-CViB-Jan14-AB B2 4.48 4.51 3.61 12.17
AL-Exome-CViB-Jan14-AB C2 4.57
AL-Exome-CViB-Jan14-AB D2 4.47

1/10 AL-Exome-CViB-Jan14-AB

Well / Set C(t) Ave. Difference nM(X10DF.)
PhiX A1 8.55 8.7
PhiX B1 8.96
PhiX C1 8.74
PhiX D1 8.56
1/10 AL-Exome-CViB-Jan14-AB G2 8.13 7.99 0.71 16.39
1/10 AL-Exome-CViB-Jan14-AB H2 7.85

1/10 AL-Exome-CViB-Jan14-CDE

Well / Set C(t) Ave. Difference nM (X10DF.)
PhiX A1 7.66 7.78
PhiX B1 8
PhiX C1 7.86
PhiX D1 7.61
1/10 AL-Exome-CViB-Jan14-CDE E3 6.57 6.68 1.1 21.44
1/10 AL-Exome-CViB-Jan14-CDE F3 6.69
1/10 AL-Exome-CViB-Jan14-CDE G3 6.72
1/10 AL-Exome-CViB-Jan14-CDE H3 6.75

AL-Exome-CViB-Jan14-CDE

Well / Set C(t) Ave. Difference nM
PhiX A1 7.22 7.32
PhiX B1 7.49
PhiX C1 7.41
PhiX D1 7.16
AL-Exome-CViB-Jan14-CDE A3 3.91 4.14 3.19 9.09
AL-Exome-CViB-Jan14-CDE B3 4.14
AL-Exome-CViB-Jan14-CDE C3 4.25
AL-Exome-CViB-Jan14-CDE D3 4.24

Result

nM
JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 1.1
1/10 Ida
AL-Exome-CViB-Jan14-AB 12.17
1/10 AL-Exome-CViB-Jan14-AB 16.39
AL-Exome-CViB-Jan14-CDE 9.09
1/10 AL-Exome-CViB-Jan14-CDE 21.44