AlanFung:LabNotes/CTCF/2011-6-6
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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