AlanFung:LabNotes/CTCF/2011-6-6: Difference between revisions
Jump to navigation
Jump to search
>Alan6017518 |
>Alan6017518 |
||
Line 32: | Line 32: | ||
95C 2m | 95C 2m | ||
95C 30s | 95C 30s | ||
52C 30s | |||
72C | 72C 2m | ||
Plate Read | Plate Read | ||
Go to step 2 repeat for 29 times | Go to step 2 repeat for 29 times | ||
72C 10m | 72C 10m | ||
15C hold | 15C hold | ||
==Results== | ==Results== |
Revision as of 20:38, 6 June 2011
Perform bisulfite conversion on 200ng of gDNA
Protocol
Setup PCR Reaction
- Add the components on ice in order while mixing gently.
Component | Amount per reaction | x6.6 |
Distilled water | 31 | 204.6 |
10X pfuTurbo Cx reaction buffer | 5 | 33 |
dNTPs (10mM each DNTP) | 1 | 6.6 |
DNA template/STD | 5 | |
CHR8 F Primer (3.3uM) | 3 | |
CHR8 R Primer (3.3uM) | 3 | |
SYBR Green (50X) | 1 | 6.6 |
PfuTurbo Cx Hotstart DNA Polymerase (2.5U/ul) | 1 | 6.6 |
Total reaction volume | 50 | 257.4 |
- Add 39ul master mix to primers and templates
- Perform Manufacturer protocol for PfuTurbo Cx Hotstart DNA Polymerase
95C 2m 95C 30s 52C 30s 72C 2m Plate Read Go to step 2 repeat for 29 times 72C 10m 15C hold