AlanFung:LabNotes/CTCF/2011-5-10

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Revision as of 21:21, 24 May 2011 by >Alan6017518 (Created page with '==Bisulfite Conversion DNA Damage Estimation== *Perform PCR with CHR8, CHR21 and CHR22 * These will be our standards File:ZhangLab_2 2011-05-10 10hr 40min.jpg * Need to purif…')
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Bisulfite Conversion DNA Damage Estimation

  • Perform PCR with CHR8, CHR21 and CHR22
  • These will be our standards

File:ZhangLab 2 2011-05-10 10hr 40min.jpg

  • Need to purify CHR8, CHR21, and CHR22.
    • Then need to estimate the concentration
    • Need amplicon length (~200)
  • Run an experiment where we do PCR on both Bisulfite-converted DNA and the three controls
    • Want to make sure number of starting molecules is the same.
  • With normal DNA, 5 pg ~= 2 molecules
  • With PCR product, X pg ~= 2 molecules; X << 5
  • Need to run a PCR reaction using 500 pg of bisulfite converted DNA and X pg of PCR product as templates, with X such that the number of molecules in each tube is the same.
    • Can run QPCR to check for DNA damage; only hot-start for 30 seconds instead of 3 minutes.