AlanFung:LabNotes/CTCF/2010-1-15: Difference between revisions
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*Only 50ng,10ng, 5ng was amplified based on the QPCR curve, will run a gel to quantify |
Revision as of 17:53, 19 January 2010
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 |
- Only 50ng,10ng, 5ng was amplified based on the QPCR curve, will run a gel to quantify