---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2017-12-11 23:05:25+00:00
layout: post
slug: dna-isolation-quantification-crassotrea-virginica-mantle-gdna
title: DNA Isolation & Quantification - Crassostrea virginica Mantle gDNA
categories:
  - 2017
  - Miscellaneous
tags:
  - Crassostrea virginica
  - DNA Isolation
  - DNA Quantification
  - E.Z.N.A. Mollusc Kit
  - Eastern oyster
  - Qubit 3.0
  - Qubit dsDNA BR
---

DNA was isolated from a single adult Eastern oyster (_Crassostrea virginica_) for a pilot project with Qiagen to test their new DNA bisulfite conversion kit. The oyster was obtained yesterday afternoon (20171210) from the [Taylo rShellfish Pioneer Square location](https://www.taylorshellfishfarms.com/location/pioneer-square). The oyster was stored @ 4C O/N.

The oyster was shucked and four pieces of upper mantle tissue (~35mg each) were snap frozen in liquid nitrogen (LN2). Tissues were pulverized under LN2 and then DNA was isolated separately from each sample using the E.Z.N.A. Mollusc DNA Kit (Omega) according to the manufcaturer's protocol.

Samples were eluted with 100uL of Elution Buffer and were pooled into a single tube.

The gDNA was quantified using the Qubit 3.0 (Invitrogen) and Qubit dsDNA Broad Range Kit (Invitrogen), using 5uL of sample.

Results:

Qubit (Google Sheet): [20171211_qubit_virginica_DNA](https://docs.google.com/spreadsheets/d/1C35zyjc10GNtBt-7z_BBr0Z_ArCZx91H5-Mc05C4evg/edit?usp=sharing)

Concentration is 58.4ng/uL.

That makes the total yield ~23.36ug (23360ng). This is more than enough to perform two separate MeDIP preps and two separate reduced representation digestions with MspI.

Will proceed with shearing of DNA for MeDIP.