AlanFung:LabNotes/CTCF/2011-6-6

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Revision as of 20:52, 6 June 2011 by >Alan6017518 (→‎Setup PCR Reaction)
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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
distilled water 34
10x pfuturbo cx reaction buffer 5
dNTPs(10mM each DNTP) 1
DNA template 2
CHR8 F primer 3.3uM 3
CHR8 R primer 3.3um 3
sybr green 50x 1
pfuturbo cx hotstart dna polymerase 2.5U/ul 1
total reaction volume 50
  • 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

Results