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Quantify libraries with Phi-X ladder qPCR[edit]
Stock Phi-X (1nM),Stock Phi-X (0.1nM), Stock Phi-X (0.05nM), JD-cpg97k+30k-Breast_cancer-01230B09-Jan15,1/10 JD-cpg97k+30k-Breast_cancer-01230B09-Jan15, 1/10 AL-Exome-CViB-Jan14-AB, 1/100 AL-Exome-CViB-Jan14-AB, 1/10 AL-Exome-CViB-Jan14-CDE, 1/100 AL-Exome-CViB-Jan14-CDE[edit]
Setup | ||
Stock Phi-X (1nM) | 1/10 JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 | 1/100 AL-Exome-CViB-Jan14-CDE |
Stock Phi-X (1nM) | 1/10JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 | 1/100 AL-Exome-CViB-Jan14-CDE |
Stock Phi-X (0.1nM) | 1/10 AL-Exome-CViB-Jan14-AB | |
Stock Phi-X (0.1nM) | 1/10 AL-Exome-CViB-Jan14-AB | |
Stock Phi-X (0.05nM) | 1/100 AL-Exome-CViB-Jan14-AB | |
Stock Phi-X (0.05nM) | 1/100 AL-Exome-CViB-Jan14-AB | |
For 2 wells | X10 | |
DNA | 2.000 | |
Syb_FP5 (100uM) | 0.200 | 2.000 |
Syb_RP7 (100uM) | 0.200 | 2.000 |
2X Phusion | 25.000 | 250.000 |
50X SYBR Green I | 0.200 | 2.000 |
H20 | 24.000 | 240.000 |
* 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_4.pdf Media:AL-Exome-CViB-Jan14-AB_4.pdf Media:AL-Exome-CViB-Jan14-CDE_4.pdf
nM | ||
0.6215000 | 0.0165400 | |
0.6598000 | 0.0178900 | |
0.9573000 | ||
1.3480000 | ||
0.0010940 | ||
0.0011790 | ||
1.5690000 | 1.7510000 | |
1.7850000 | 1.3480000 | |
Average (nM) | ||
0.641 | 0.017 | |
1.153 | ||
0.001 | ||
1.677 | 1.550 | |
Result[edit]
X DF
nM | ||
JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 | 1.677 | |
1/10JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 | 6.41 | |
1/10 AL-Exome-CViB-Jan14-AB | 11.530 | |
1/100 AL-Exome-CViB-Jan14-AB | 0.110 | |
1/10 AL-Exome-CViB-Jan14-CDE | 15.5 | |
1/100 AL-Exome-CViB-Jan14-CDE | 1.72 | |