---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2010-07-03 04:46:01+00:00
layout: post
slug: medip-sbwb-fragmented-gdna-continued-from-yesterday
title: MeDIP - SB/WB Fragmented gDNA (continued from yesterday)
categories:
  - 2010
  - Miscellaneous
tags:
  - Crassostrea gigas
  - EtOH precipitation
  - MeDIP
  - Pacific oyster
---

Continued MeDIP process from yesterday. Protein A/G beads were pelleted XXXXXXXXX, supe transferred to clean tube. Beads were washed 3x in the following fashion, each wash saved to retain unmethylated DNA:

1.

Samples were phenol:chloroform extracted and EtOH precipitated:





  1. Added equal volume of phenol:chloroform:IAA, vortexed, spun @ 12,500g, 5mins, 4C.



  2. Transferred aqueous phase to clean tube.



  3. Added equal volume of chloroform, vortexed, spun @ 12,500g, 5mins, 4C.



  4. Transferred aqueous phase to clean tube.



  5. Added 0.1 vols 3M NaOAc (pH=5.2), 2.5 vols of 100% EtOH, mixed and stored @ -20C over the weekend.






Will finish precipitation next week and quantify recovery.
