---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2018-01-10 21:33:50+00:00
layout: post
slug: mbd-enrichment-crassostrea-virginica-sheared-dna-day-3
title: MBD Enrichment – Crassostrea virginica Sheared DNA Day 3
categories:
  - 2018
  - Miscellaneous
tags:
  - Crassostrea virginica
  - Eastern oyster
  - EtOH precipitation
  - MBD
  - MBD enrichment
  - MethylMiner Methylated DNA Enrichment Kit
---

Continued MBD enrichment of _C.virginica_ DNA [from yesterday](https://robertslab.github.io/sams-notebook/posts/2018/2018-01-09-mbd-enrichment…ared-dna-day-2/) for Qiagen project.

Followed the MethylMiner Methylated DNA Enrichment Kit (Invitrogen) manufacturer's protocol for input DNA amounts of 1 -10ug (I am using 8ug in each of two samples).

Since the protocol has two elution steps that are each saved separately from each other for each sample, I did the following to combine the two elution fractions into a single sample:




    
  * Pelleted one elution fraction from each sample

    
  * Discarded supernatant from pelleted sample

    
  * Transferred second elution fraction to the pellet from the first elution fraction

    
  * Pelleted second elution fraction



The rest of the ethanol precipitation procedure was followed per the manufacturer's protocol.

Final pellets were resuspended in 25μL of Buffer EB (Qiagen) and stored temporarily on ice for quantification.
