---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2015-09-02 00:05:22+00:00
layout: post
slug: genomic-dna-isolation-olympia-oyster-adductor-musle-mantle
title: Genomic DNA Isolation - Olympia oyster adductor musle & mantle
categories:
  - 2015
  - Olympia Oyster Genome Sequencing
tags:
  - DNA Isolation
  - DNA Quantification
  - E.Z.N.A. Mollusc Kit
  - gDNA
  - NanoDrop1000
  - olympia oyster
  - Ostrea lurida
---

Isolated gDNA from _Ostrea lurida_ (Olympia oyster) [adductor muscle & mantle samples collected by Brent & Steven on 20150812](https://onsnetwork.org/halfshell/2015/08/12/another-day-another-species/) using the E.Z.N.A Mollusc DNA Kit (Omega Bio-Tek) according to the manufacturer’s protocol, with the following adjustments:




    
  * 29.8mg of adductor muscle

    
  * 28.7mg of mantle

    
  * Tissues homogenized in 350μL of ML1 Buffer with disposable mortar/pestle tubes using only three pestles strokes

    
  * Homogenized tissue incubated in ML1 Buffer + Proteinase K @ 60C for 2.5hrs

    
  * Added 310μL of MBL Buffer to adductor muscle sample and 265μL of MBL Buffer to mantle sample

    
  * Added 620μL of 100% EtOH and 530μL of 100% EtOH to the adductor muscle and mantle sample, respectively.

    
  * Eluted with 50μL Elution Buffer.



Spec’d on Roberts Lab NanoDrop1000 (ThermoFisher) and stored temporarily at 4C to avoid freeze-thawing before sending off for sequencing.



Results:



![](https://eagle.fish.washington.edu/Arabidopsis/20150901_gDNA_oly_ODs.JPG)(http://eagle.fish.washington.edu/Arabidopsis/20150901_gDNA_oly_ODs.JPG)



![](https://eagle.fish.washington.edu/Arabidopsis/20150901_gDNA_oly_plots.JPG)(http://eagle.fish.washington.edu/Arabidopsis/20150901_gDNA_oly_plots.JPG)



Yields are good (~6μg for the adductor muscle and ~10μg for the mantle).

The adductor muscle sample looks pretty good (perfect 260/280 ratio and solid 260/230 ratio), while the mantle sample looks OK (good 260/280 ratio, tolerable 260/230 ratio). Will run samples on gel to assess gDNA integrity.


