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>Alan6017518 (Created page with '==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== {| {{table}} | Setup|||| |- | 1…') |
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==Quantify libraries with Phi-X ladder qPCR== | ==Quantify libraries with Phi-X ladder qPCR== | ||
==AL-Exome-CViB-Jan14-AB | ==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}} | {| {{table}} | ||
| Setup|||| | | 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}} | {| {{table}} | ||
| ||2 | | ||For 2 wells||X10 | ||
|- | |- | ||
| DNA||2|| | | DNA||2.000|| | ||
|- | |- | ||
| Syb_FP5 (100uM)||0. | | Syb_FP5 (100uM)||0.200||2.000 | ||
|- | |- | ||
| Syb_RP7 (100uM)||0. | | Syb_RP7 (100uM)||0.200||2.000 | ||
|- | |- | ||
| 2X Phusion||25|| | | 2X Phusion||25.000||250.000 | ||
|- | |- | ||
| 50X SYBR Green||0. | | 50X SYBR Green I||0.200||2.000 | ||
|- | |- | ||
| | | H20||24.000||240.000 | ||
|- | |- | ||
| | | | ||
|} | |} | ||
* Dilute sample, 1ul in 9ul ddh2o | * Dilute sample, 1ul in 9ul ddh2o | ||
* mix 2ul 1/10 sample with 98ul mix in non-stick tube | * mix 2ul 1/10 sample with 98ul mix in non-stick tube | ||
Line 48: | Line 40: | ||
98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min | 98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min | ||
[[Media:JD-cpg97k+30k-Breast_cancer-01230B09- | [[Media:JD-cpg97k+30k-Breast_cancer-01230B09-Jan15_log_4.pdf]] | ||
[[Media:AL-Exome-CViB-Jan14- | [[Media:AL-Exome-CViB-Jan14-AB_4.pdf]] | ||
[[Media:AL-Exome-CViB-Jan14- | [[Media:AL-Exome-CViB-Jan14-CDE_4.pdf]] | ||
{| {{table}} | {| {{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== | ==Result== | ||
X DF | |||
{| {{table}} | {| {{table}} | ||
| ||||nM | | ||||nM | ||
|- | |- | ||
| JD-cpg97k+30k-Breast_cancer-01230B09-Jan15||||1. | | JD-cpg97k+30k-Breast_cancer-01230B09-Jan15||||1.677 | ||
|- | |- | ||
| 1/ | | 1/10JD-cpg97k+30k-Breast_cancer-01230B09-Jan15||||6.41 | ||
|- | |||
| |||| | |||
|- | |- | ||
| AL-Exome-CViB-Jan14-AB|||| | | 1/10 AL-Exome-CViB-Jan14-AB||||11.530 | ||
|- | |- | ||
| 1/ | | 1/100 AL-Exome-CViB-Jan14-AB||||0.110 | ||
|- | |- | ||
| |||| | | |||| | ||
|- | |- | ||
| AL-Exome-CViB-Jan14-CDE|||| | | 1/10 AL-Exome-CViB-Jan14-CDE||||15.5 | ||
|- | |- | ||
| 1/ | | 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 | |