Revision as of 17:51, 19 January 2010 by >Alan6017518
Contents
- 1 Quantify Bisulfite Converted Samples with Qubit ssDNA Kit
- 2 On 50ng, 10ng, 5ng, 1ng, 0.5ng, 0.1ng & 0.05ng P21-200BP
- 3 PCR quantification of Bisulfite treated samples using 3 paris of CHR bilsulfite conversion primers
- 4 On 50ng, 10ng, 5ng, 1ng, 0.5ng, 0.1ng & 0.05ng P21-200BP
Quantify Bisulfite Converted Samples with Qubit ssDNA Kit
On 50ng, 10ng, 5ng, 1ng, 0.5ng, 0.1ng & 0.05ng P21-200BP
7 samples
Prepare 7 Qubit Assay tubes
Prepare 7*200ul diluted dye=1400ul
Mix 7ul dye with 1393ul buffer
Mix 2ul samples with 98ul diluted dye
vortex spin down and wait for 2 mins
Result
Start (ng) |
Sample Concentration ng/ul |
Volume |
Amount Recovered (ng) |
Yield %
|
50 |
1.7 |
20 |
34 |
68
|
10 |
0.25 |
20 |
5 |
50
|
5 |
0.13 |
20 |
2.6 |
52
|
1 |
0.26 |
20 |
5.2 |
520
|
0.5 |
N/A |
20 |
|
|
0.1 |
N/A |
20 |
|
|
0.05 |
N/A |
20 |
|
|
|
PCR quantification of Bisulfite treated samples using 3 paris of CHR bilsulfite conversion primers
On 50ng, 10ng, 5ng, 1ng, 0.5ng, 0.1ng & 0.05ng P21-200BP
1 sample |
CHR22 |
CHR21 |
CHR8
|
2X iQ Super Mix |
20 |
20 |
20
|
Primer F (3.3uM) |
6 |
6 |
6
|
Primer R (3.3uM) |
6 |
6 |
6
|
|
|
|
|
7 Samples |
CHR22 |
CHR21 |
CHR8
|
2X iQ Super Mix |
160 |
160 |
160
|
Primer F (3.3uM) |
48 |
48 |
48
|
Primer R (3.3uM) |
48 |
48 |
48
|
|
|
|
|
|
Setup |
|
|
|
CHR22 |
CHR21 |
CHR8
|
1 |
50ng |
50ng |
50ng
|
2 |
10ng |
10ng |
10ng
|
3 |
5ng |
5ng |
5ng
|
4 |
1ng |
1ng |
1ng
|
5 |
0.5ng |
0.5ng |
0.5ng
|
6 |
0.1ng |
0.1ng |
0.1ng
|
7 |
0.05ng |
0.05ng |
0.05ng
|
|