AlanFung:LabNotes/CTCF/2011-6-28

From ZhangLabWiki
Revision as of 00:02, 28 June 2011 by >Alan6017518 (→‎Proteinase K Digestion)
Jump to navigation Jump to search

MDA Bisulfite Converted Sequencing Library Consturction

Objective

  • Work on sperm/ESCs control cell samples plus GM12878 gDNA

Overview

  • Proteinase K Digestion
  • Bisulfite conversion
  • MDA
  • EtOH precipitation
  • Nextera Library Construction (Low input protocol)
  • Page Analysis
  • Sanger Sequencing

Protocol

Proteinase K Digestion

  1. Sperms: ~50cell/tube; 15 tubes
  2. ES cells: ~50cell/tube; 15 tubes
  • Thaw 4 tubes on ice, we will use 100cells per reaction

Bisulfite Conversion

MDA

Ethanol Precipitation

Nextera Library Construction (Low input protocol)

PAGE Analysis

Sanger Analysis