---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2015-08-29 00:15:02+00:00
layout: post
slug: genomic-dna-isolation-geoduck-adductor-muscle-foot
title: Genomic DNA Isolation - Geoduck Adductor Muscle & Foot
categories:
  - 2015
  - Geoduck Genome Sequencing
tags:
  - adductor muscle
  - DNA Isolation
  - DNA Quantification
  - E.Z.N.A. Mollusc Kit
  - foot
  - gDNA
  - geoduck
  - NanoDrop1000
  - Panopea generosa
---

Isolated gDNA from _Panopea generosa_ (geoduck) [adductor muscle & foot samples collected by Brent & Steven on 20150811](https://onsnetwork.org/halfshell/2015/08/11/big-day-big-clam/) using the E.Z.N.A Mollusc DNA Kit (Omega Bio-Tek) according to the manufacturer's protocol, with the following adjustments:




    
  * 41.8mg of adductor muscle

    
  * 30.0mg of foot used

    
  * 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 265μL of MBL Buffer

    
  * Added 514μL of 100% EtOH.

    
  * Eluted with 75μL Elution Buffer.



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



Results:



![](https://eagle.fish.washington.edu/Arabidopsis/20150828_geoduck_gDNA_ODs.JPG)(http://eagle.fish.washington.edu/Arabidopsis/20150828_geoduck_gDNA_ODs.JPG)



![](https://eagle.fish.washington.edu/Arabidopsis/20150828_geoduck_gDNA_plots.JPG)(http://eagle.fish.washington.edu/Arabidopsis/20150828_geoduck_gDNA_plots.JPG)



Yields are good (~7ug).

Quality (260/280 ratios) looks great for both samples.

260/230 ratio not very good for adductor muscle, but perfect for foot tissue.

Will run samples on gel to assess gDNA integrity.


