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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 | ||
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 |
2 Wells | X13 | |
DNA | 2 | 26 |
Syb_FP5 (100uM) | 0.2 | 2.6 |
Syb_RP7 (100uM) | 0.2 | 2.6 |
2X Phusion | 25 | , |
50X SYBR Green | 0.2 | 2.6 |
H2O | 24 | 312 |
Total | 345.8 | |
* 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
C(t) | @0.025 | Average | |||||
PhiX | A1 | 8.830 | 8.826 | 8.630 | |||
PhiX | B1 | 8.630 | 8.634 | ||||
PhiX | D1 | 8.430 | 8.429 | ||||
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 | ||||
Ida | H1 | 8.280 | 8.282 | ||||
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 | ||||
1/10 AL-Exome-CViB-Jan14-AB | H2 | 7.020 | 7.017 | ||||
C(t) | @0.015 | Average | |||||
PhiX | A1 | 7.640 | 7.642 | 7.505 | |||
PhiX | B1 | 7.520 | 7.517 | ||||
PhiX | D1 | 7.350 | 7.355 | ||||
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 | ||||
C(t) | Average | ||||||
PhiX | A1 | 7.640 | 7.5 | ||||
PhiX | B1 | 7.520 | |||||
PhiX | D1 | 7.350 | |||||
AL-Exome-CViB-Jan14-CDE | A3 | 4.050 | Average | Difference | nM | ||
AL-Exome-CViB-Jan14-CDE | B3 | 3.810 | 3.99 | 3.51 | 11.42 | ||
AL-Exome-CViB-Jan14-CDE | C3 | 4.080 | |||||
AL-Exome-CViB-Jan14-CDE | D3 | 4.020 | |||||
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 | |
1/10 AL-Exome-CViB-Jan14-CDE | G3 | 5.850 | |||||
1/10 AL-Exome-CViB-Jan14-CDE | H3 | 5.680 | |||||