Revision as of 21:43, 21 January 2010 by >Alan6017518
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 |
|
|
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
|
|