AlanFung:LabNotes/CTCF/2011-4-25
Jump to navigation
Jump to search
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
- Improve the yield from the bisulfite conversion step using suggestions made by zymo tech support
- Increase yield from the bisulfite conversion using zymo methylation direct kit (works with 50pg starting amount)
- Test Kapa Hifi polymerase
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 (Methylation Direct Kit)