Revision as of 22:20, 19 November 2009 by >Alan6017518
Quantification of New amplified Phi-X and all libraries[edit]
- Dilute all samples by 10 fold
Samples |
|
|
|
|
PhiX-Stock |
CVF |
PGP4 |
PGP10 |
PGP8C
|
PhiX-Stock |
CVF |
PGP4 |
PGP10 |
PGP8C
|
PhiX-Stock |
1-7 PGP1 |
PGP6 |
NA12878 |
PGP8D
|
PhiX-Stock |
1-7 PGP1 |
PGP6 |
NA12878 |
PGP8D
|
PhiX |
8 PGP1 |
PGP8 |
PGP8A |
PGP8E
|
PhiX |
8 PGP1 |
PGP8 |
PGP8A |
PGP8E
|
CVI |
9 PGP1 |
PGP9 |
PGP8B |
N12
|
CVI |
9 PGP1 |
PGP9 |
PGP8B |
N12
|
|
- split every 50ul rxn into 2 wells
98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min
|
|
X20 |
X1.1
|
PhiX Control DNA (10nM) |
1.000 |
|
|
Syb_FP5 (100uM) |
0.200 |
4.000 |
4.400
|
Syb_RP7 (100uM) |
0.200 |
4.000 |
4.400
|
2X Phusion |
25.000 |
500.000 |
550.000
|
50X SYBR Green I |
0.200 |
4.000 |
4.400
|
H20 |
24.000 |
480.000 |
528.000
|
|
Results[edit]
Result |
|
|
|
|
PhiX |
9.112 |
Cycle Difference |
2^ |
x10 (nM)
|
Amp PhiX |
6.051 |
3.062 |
8.348 |
83.484
|
CVI |
6.877 |
2.236 |
4.709 |
47.093
|
CVF |
8.155 |
0.958 |
1.942 |
19.419
|
1-7 PGP1 |
8.346 |
0.766 |
1.701 |
17.011
|
8 PGP1 |
6.510 |
2.603 |
6.073 |
60.734
|
9 PGP1 |
7.244 |
1.868 |
3.650 |
36.503
|
N12 |
7.915 |
1.198 |
2.293 |
22.934
|
PGP8B |
11.776 |
2.664 |
6.339 |
1.577
|
|
Dilution[edit]
Dilute to 10nM |
Sample (ul) |
ddH2O |
|
Amp PhiX |
5.000 |
36.742 |
|
CVI |
5.000 |
18.546 |
|
CVF |
5.000 |
4.710 |
|
1-7 PGP1 |
5.000 |
3.506 |
|
8 PGP1 |
5.000 |
25.367 |
|
9 PGP1 |
5.000 |
13.251 |
|
N12 |
5.000 |
6.467 |
|
PGP8B |
5.000 |
2.885 |
1nM
|
|