Revision as of 21:41, 21 January 2010 by >Alan6017518
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
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
JD-cpg97k+30k-Breast_cancer-01230B09-Jan15
|
Well / Set |
C(t) |
|
Difference |
nM
|
PhiX |
A1 |
6.98 |
7.06 |
|
|
PhiX |
B1 |
7.2 |
|
|
|
PhiX |
C1 |
7.16 |
|
|
|
PhiX |
D1 |
6.89 |
|
|
|
JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 |
E1 |
6.82 |
6.92 |
0.14 |
1.1
|
JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 |
F1 |
7.09 |
|
|
|
JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 |
G1 |
6.89 |
|
|
|
JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 |
H1 |
6.86 |
|
|
|
|
AL-Exome-CViB-Jan14-AB
|
Well / Set |
C(t) |
Ave. |
Difference |
nM
|
|
|
|
|
|
|
|
|
|
|
|
|
PhiX |
A1 |
7.99 |
8.11 |
|
|
PhiX |
B1 |
8.33 |
|
|
|
PhiX |
C1 |
8.18 |
|
|
|
PhiX |
D1 |
7.95 |
|
|
|
AL-Exome-CViB-Jan14-AB |
B2 |
4.48 |
4.51 |
3.61 |
12.17
|
AL-Exome-CViB-Jan14-AB |
C2 |
4.57 |
|
|
|
AL-Exome-CViB-Jan14-AB |
D2 |
4.47 |
|
|
|
|
1/10 AL-Exome-CViB-Jan14-AB
|
Well / Set |
C(t) |
Ave. |
Difference |
nM(X10DF.)
|
PhiX |
A1 |
8.55 |
8.7 |
|
|
PhiX |
B1 |
8.96 |
|
|
|
PhiX |
C1 |
8.74 |
|
|
|
PhiX |
D1 |
8.56 |
|
|
|
1/10 AL-Exome-CViB-Jan14-AB |
G2 |
8.13 |
7.99 |
0.71 |
16.39
|
1/10 AL-Exome-CViB-Jan14-AB |
H2 |
7.85
|
|
1/10 AL-Exome-CViB-Jan14-CDE
|
Well / Set |
C(t) |
Ave. |
Difference |
nM (X10DF.)
|
PhiX |
A1 |
7.66 |
7.78 |
|
|
PhiX |
B1 |
8 |
|
|
|
PhiX |
C1 |
7.86 |
|
|
|
PhiX |
D1 |
7.61 |
|
|
|
1/10 AL-Exome-CViB-Jan14-CDE |
E3 |
6.57 |
6.68 |
1.1 |
21.44
|
1/10 AL-Exome-CViB-Jan14-CDE |
F3 |
6.69 |
|
|
|
1/10 AL-Exome-CViB-Jan14-CDE |
G3 |
6.72 |
|
|
|
1/10 AL-Exome-CViB-Jan14-CDE |
H3 |
6.75 |
|
|
|
|
AL-Exome-CViB-Jan14-CDE
|
Well / Set |
C(t) |
Ave. |
Difference |
nM
|
PhiX |
A1 |
7.22 |
7.32 |
|
|
PhiX |
B1 |
7.49 |
|
|
|
PhiX |
C1 |
7.41 |
|
|
|
PhiX |
D1 |
7.16 |
|
|
|
AL-Exome-CViB-Jan14-CDE |
A3 |
3.91 |
4.14 |
3.19 |
9.09
|
AL-Exome-CViB-Jan14-CDE |
B3 |
4.14 |
|
|
|
AL-Exome-CViB-Jan14-CDE |
C3 |
4.25 |
|
|
|
AL-Exome-CViB-Jan14-CDE |
D3 |
4.24 |
|
|
|
|
Result
|
|
nM
|
JD-cpg97k+30k-Breast_cancer-01230B09-Jan15 |
|
1.1
|
1/10 Ida |
|
|
AL-Exome-CViB-Jan14-AB |
|
12.17
|
1/10 AL-Exome-CViB-Jan14-AB |
|
16.39
|
|
|
|
AL-Exome-CViB-Jan14-CDE |
|
9.09
|
1/10 AL-Exome-CViB-Jan14-CDE |
|
21.44
|
|