AlanFung:LabNotes/CTCF/2011-3-29

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Revision as of 16:19, 29 March 2011 by >Alan6017518 (Created page with '='''Nextera Methyl-Seq DNA Sample Prep on 100cells (600pg DNA)'''= ="""Low input transposase-based library preparation (Shendure's Protocol)'''= ==Objective== *Confirmation of …')
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Nextera Methyl-Seq DNA Sample Prep on 100cells (600pg DNA)

"""Low input transposase-based library preparation (Shendure's Protocol)

Objective

  • Confirmation of the library prep with beta nextera methyl-seq kit with input DNA (100 cells, ~600pg DNA starting amount) with GM12878 gDNA.
  • Adapt Shendure's protocol for low input library preparation
  • Perform bis-cut with carrier and replace column purification step during bis-cut with ethanol precipitation


Samples & Materials

  • Nextera Sample Prep Kit_Epicentre_NMS072910B
NexteraTM Methyl-Seq Tagmentation Mix (Illumina-compatible) 
5X Nextera Reaction Buffer (HMW)
50X Nextera Primer Cocktail (Illumina-compatible) 
50X Nextera Adaptor 2 (Illumina-compatible) 
2X Nextera Fill-in Buffer 
2X Nextera PCR Buffer 
200X Nextera Read 1 Primer 
200X Nextera Read 2 Primer 
200X Nextera Index Read Primer  
Nextera Methyl-Seq Polymerase 
  • Input DNA
  • Bisulfite conversion kit
  • Qiagen Purification Kit

Overview

  • Tagmentation
  • Purification
  • Fill-in
  • Purification
  • Bisulfite Conversion with lambda phage DNA)
  • Purification
  • bPCR-compatible sites and library enrichment