---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
layout: post
title: MBD Enrichment - Ethanol Precipitation of C.virginica MBD samples
date: '2019-03-12 09:49'
tags:
  - MBD
  - methylminer
  - precipitation
  - Crassostrea virginica
  - eastern oyster
categories:
  - 2019
  - Miscellaneous
---
After completing two rounds of MBD enrichment ([first 12 samples on 20190307](https://robertslab.github.io/sams-notebook/posts/2019/2019-03-07-MBD-Enrichment-C.virginica-Sheared-Mantle-DNA-from-20190306/) and the [second 12 samples on 20190311](https://robertslab.github.io/sams-notebook/2019/03/11/MBD-Enrichment-C.virginica-Sheared-Mantle-DNA-from-20190306/)), I performed ethanol precipitation, per the MethylMiner instructions with the following modifications:


- after additions of glycogen, sodium acetate and ethanol, incubated samples O/N at -80<sup>o</sup>C

- after incubation, the two elutions were consolidated in the following fashion:
  - the first of each of the two elutions was pelleted, supe discarded, and second elution transferred to the pellet
  - second elution was pelleted in same tube as first

- spins were performed at 13,000g

Samples were resuspended in 25uL of PCR H<sub>2</sub>O.

Currently, don't have appropriate tubes for quantification, so will do so once they arrive.

Samples were stored in the small -20<sup>o</sup>C freezer in the following box (Google Sheet):

- [Sam gDNA Box 3](https://docs.google.com/spreadsheets/d/16n4LzVNXrMd3fYRHN4UR4CQ67yRXwlJfDiAmbxD284g/edit?usp=sharing)
