---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2010-07-10 04:27:32+00:00
layout: post
slug: gdna-isolation-various-gigas-samples-continued-from-yesterday
title: gDNA Isolation - Various gigas samples (continued from yesterday)
categories:
  - 2010
  - Miscellaneous
tags:
  - 5-azacitidine
  - Crassostrea gigas
  - DNA Isolation
  - DNA Quantification
  - DNazol
  - gill
  - mantle
  - NanoDrop1000
  - Pacific oyster
  - Vibrio exposure
  - Vibrio tubiashii
---

Pelleted residual tissue 10mins @ 10,000g @ RT. Transferred supe to new tubes. Precipitated DNA with 0.25mL 100% EtOH. Incubated 3mins @ RT. DNA was pelleted 5mins @ 5000g @ RT. Supe was removed, pellets were washed with 1mL 75% EtOH (x2). Supe was fully removed and the DNAs were resuspended in 300uL 8mM NaOH (made 7/9/10 SJW).

1M HEPES (provided with DNAzol) was added at a 1:100 dilution to achieve a pH = 8.0. This was based on the [DNAzol protocol calculations](https://www.mrcgene.com/dnazol.htm) (For 1mL of 8mM NaOH, use 101uL of 0.1M HEPES = pH 8.0).

Samples were spec'd on NanoDrop 1000. Used a sample with 8mM NaOH and 1M HEPES as a blank to match the pH = 8.0 of the samples.

Results:

![](https://eagle.fish.washington.edu/Arabidopsis/20100709%20gDNA%20ODs.JPG)

260/280 ratios look good for all samples. Most of the samples have mediocre 260/230 ratios. Yields are excellent for all samples.
