Revision as of 18:18, 23 November 2009 by >Alan6017518
Quantification of New amplified Phi-X and all libraries
- Dilute concentration by 10 fold for Har_2, Har_5
- Dilute New amplified PhiX to 1nM (Mix 2ul amp. PhiX with 164.97 ddH2O)
- Keep P3, P5, P7 and CVF_N_D as is
Samples |
|
|
|
|
PhiX-Stock |
8/9 P7
|
PhiX-Stock |
8/9 P7
|
PhiX(1nM) |
Har_2
|
PhiX(1nM) |
Har_2
|
8/9 P3 |
Har_5
|
8/9 P3 |
Har_5
|
8/9 P5 |
CVF_N_D
|
8/9 P5 |
CVF_N_D
|
|
- split every 50ul rxn into 2 wells
98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min
|
|
X8 |
X1.1
|
PhiX Control DNA (10nM) |
1.000 |
|
|
Syb_FP5 (100uM) |
0.200 |
1.6 |
1.76
|
Syb_RP7 (100uM) |
0.200 |
1.6 |
1.76
|
2X Phusion |
25.000 |
200 |
220
|
50X SYBR Green I |
0.200 |
1.6 |
1.76
|
H20 |
24.000 |
192 |
211.2
|
|
|
nM |
sample ul |
Tris-CL ul |
HP3 |
total
|
8/9 P7 |
8.09 |
3 |
20.06 |
1.21 |
24.27
|
CVF_N_D |
2.91 |
4 |
7.06 |
0.58 |
11.64
|
|
Dilute to 10nM |
Sample (ul) |
|
ddH2O
|
8/9 P3 |
4.000 |
6.928 |
2.928
|
8/9 P5 |
4.000 |
6.628 |
2.628
|
Har_2 |
4.000 |
5.600 |
1.600
|
Har_5 |
4.000 |
6.876 |
2.876
|
|