AlanFung:LabNotes/CTCF/2011-2-14
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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