4th: Difference between revisions

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>Alan6017518
(/* 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)
>Alan6017518
(/* 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)
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    * 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

Revision as of 18:59, 21 January 2010

Quantify libraries with Phi-X ladder qPCR

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

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