Revision as of 22:42, 21 January 2010 by >Alan6017518
Quantify libraries with Phi-X ladder qPCR[edit]
AL-Exome-CViB-Jan14-AB w/ 1/10 dilution, AL-Exome-CViB-Jan14-CDE w/ 1/10 dilution, Ida's library[edit]
Setup |
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|
1nm PhiX |
AL-Exome-CViB-Jan14-AB |
AL-Exome-CViB-Jan14-CDE
|
1nm PhiX |
AL-Exome-CViB-Jan14-AB |
AL-Exome-CViB-Jan14-CDE
|
1nm PhiX |
AL-Exome-CViB-Jan14-AB |
AL-Exome-CViB-Jan14-CDE
|
1nm PhiX |
AL-Exome-CViB-Jan14-AB |
AL-Exome-CViB-Jan14-CDE
|
Ida |
1/10 AL-Exome-CViB-Jan14-AB |
1/10 AL-Exome-CViB-Jan14-CDE
|
Ida |
1/10 AL-Exome-CViB-Jan14-AB |
1/10 AL-Exome-CViB-Jan14-CDE
|
Ida |
1/10 AL-Exome-CViB-Jan14-AB |
1/10 AL-Exome-CViB-Jan14-CDE
|
Ida |
1/10 AL-Exome-CViB-Jan14-AB |
1/10 AL-Exome-CViB-Jan14-CDE
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2 Wells |
X13
|
DNA |
2 |
26
|
Syb_FP5 (100uM) |
0.2 |
2.6
|
Syb_RP7 (100uM) |
0.2 |
2.6
|
2X Phusion |
25 |
,
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50X SYBR Green |
0.2 |
2.6
|
H2O |
24 |
312
|
Total |
|
345.8
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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
Media:JD-cpg97k+30k-Breast_cancer-01230B09-Jan15_log.pdf
Media:AL-Exome-CViB-Jan14-AB.pdf
Media:AL-Exome-CViB-Jan14-CDE.pdf
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C(t) |
@0.025 |
Average |
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PhiX |
A1 |
8.830 |
8.826 |
8.630 |
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PhiX |
B1 |
8.630 |
8.634 |
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PhiX |
D1 |
8.430 |
8.429 |
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Ida |
E1 |
8.260 |
8.257 |
Average |
Difference |
nM |
|
Ida |
F1 |
8.500 |
8.496 |
8.277 |
0.353 |
1.277 |
|
Ida |
G1 |
8.070 |
8.073 |
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Ida |
H1 |
8.280 |
8.282 |
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1/10 AL-Exome-CViB-Jan14-AB |
E2 |
7.290 |
7.289 |
Average |
Difference |
nM |
X10
|
1/10 AL-Exome-CViB-Jan14-AB |
F2 |
7.430 |
7.433 |
7.173 |
1.457 |
2.745 |
27.45
|
1/10 AL-Exome-CViB-Jan14-AB |
G2 |
6.950 |
6.953 |
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1/10 AL-Exome-CViB-Jan14-AB |
H2 |
7.020 |
7.017 |
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C(t) |
@0.015 |
Average |
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|
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PhiX |
A1 |
7.640 |
7.642 |
7.505 |
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|
PhiX |
B1 |
7.520 |
7.517 |
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PhiX |
D1 |
7.350 |
7.355 |
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AL-Exome-CViB-Jan14-AB |
A2 |
4.290 |
4.292 |
Average |
Difference |
nM |
|
AL-Exome-CViB-Jan14-AB |
B2 |
4.260 |
4.261 |
4.306 |
3.199 |
9.181 |
|
AL-Exome-CViB-Jan14-AB |
C2 |
4.360 |
4.365 |
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C(t) |
Average |
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|
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PhiX |
A1 |
7.640 |
7.5 |
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PhiX |
B1 |
7.520 |
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PhiX |
D1 |
7.350 |
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AL-Exome-CViB-Jan14-CDE |
A3 |
4.050 |
Average |
Difference |
nM |
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AL-Exome-CViB-Jan14-CDE |
B3 |
3.810 |
3.99 |
3.51 |
11.42 |
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AL-Exome-CViB-Jan14-CDE |
C3 |
4.080 |
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AL-Exome-CViB-Jan14-CDE |
D3 |
4.020 |
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1/10 AL-Exome-CViB-Jan14-CDE |
E3 |
6.000 |
Average |
Difference |
nM |
X10 |
|
1/10 AL-Exome-CViB-Jan14-CDE |
F3 |
6.130 |
5.92 |
1.59 |
3.01 |
30.070 |
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1/10 AL-Exome-CViB-Jan14-CDE |
G3 |
5.850 |
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1/10 AL-Exome-CViB-Jan14-CDE |
H3 |
5.680 |
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