AlanFung:LabNotes/CTCF/2011-6-30: Difference between revisions

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*Repeat 6/28 experiment with only gDNA (UV EB buffer, use RT-PCR grade water as NTC)
*Repeat 6/28 experiment with only gDNA (UV EB buffer, use RT-PCR grade water as NTC)
*Sample List
*Sample List
#A-gDNA W 0.1ul proteinase K
#A-600pg gDNA W 0.1ul proteinase K
#B-gDNA W proteinase k
#B-600pg gDNA W proteinase k
#C-gDNA W/O proteinase k
#C-600pg gDNA W/O proteinase k
#D-gDNA W protease
#D-600pg gDNA W protease
#E-NTC W proteinase K
#E-NTC W proteinase K
#F-NTC W/O proteinase K
#F-NTC W/O proteinase K

Revision as of 18:56, 30 June 2011

N9-MDA Bisulfite Converted Sequencing Library Construction

Objective

  • Figure out source of contamination from 6/28 experiment
  • Repeat 6/28 experiment with only gDNA (UV EB buffer, use RT-PCR grade water as NTC)
  • Sample List
  1. A-600pg gDNA W 0.1ul proteinase K
  2. B-600pg gDNA W proteinase k
  3. C-600pg gDNA W/O proteinase k
  4. D-600pg gDNA W protease
  5. E-NTC W proteinase K
  6. F-NTC W/O proteinase K

Overview

  1. Proteinase K/Protease Digestion
  2. Bisulfite Conversion
  3. Heat Denature
  4. MDA
  5. Ethanol Precipitation
  6. Nextera Library Construction (Low input protocol)
  7. PAGE Analysis

Protocol

Proteinase K/Protease Digestion