Home Protocols Elute Captured Methylated CpG DNA (E2600)

Elute Captured Methylated CpG DNA (E2600)

Introduction

For all downstream techniques, including endpoint and real-time PCR assays, bisulfite conversion, cloning and sequencing, direct sequencing, library preparation for high-throughput sequencing, labeling for DNA microarray analysis or methylation-sensitive restriction enzyme-based assays, heating the sample at 65°C for 15 minutes is the recommended method of choice for elution of DNA. The appendix section also includes a protocol for NaCl multi-fraction elution series, followed by Monarch spin column cleanup if necessary.

Protocol

Heat Method

  1. Add 50–100 µl of DNase-free water to the sample of magnetic beads pellet. Mix beads by flicking the tube. Incubate the slurry in a heat block or thermomixer set at 65°C for 15 minutes, with frequent mixing.
  2. Briefly centrifuge the sample.
  3. Place the tube on the magnetic rack for 2–5 minutes to concentrate the beads on the wall of the tube.
  4. Carefully remove the supernatant to a fresh microcentrifuge tube. This eluted supernatant contains enriched methyl CpG containing DNA. Proceed directly to downstream analysis or store the eluted DNA sample at –20°C.