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==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==
{| {{table}}
| 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||
|-
|
|}
{| {{table}}
| ||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]]
{| {{table}}
| 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==
X DF
{| {{table}}
| ||||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
|-
|
|}

Latest revision as of 21:43, 21 January 2010

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