AlanFung:LabNotes/CTCF/2011-2-14

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Revision as of 01:35, 15 February 2011 by >Alan6017518 (→‎Objective)
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Nextera Methyl-Seq DNA Sample Prep

Objective

  • Confirmation of the library prep with beta nextera methyl-seq kit with input DNA (100ng starting amount) and size around 200-1000BP, by performing tagmentation on the input DNA and running a gel will tell wether we can make libraries with the chip DNA.

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
  • Purification
  • bPCR-compatible sites and library enrichment

Protocol

  • Nuclease Free water
100ng input DNA 
4ul 5x Nextera HMW Reaction Buffer
1ul Nextera Methyl-Seq Tagmentation Mix

Results

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