---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2009-05-13 17:10:43+00:00
layout: post
slug: gdna-isolation-c-pugetti
title: gDNA Isolation - C.pugetti
categories:
  - 2009
  - Miscellaneous
tags:
  - Cycloclasticus pugetii
  - DNA Isolation
  - gDNA
  - gel
  - JGI
---

Isolated according to [JGI protocol](https://my.jgi.doe.gov/general/protocols/DNA_Isolation_Bacterial_CTAB_Protocol.doc) (Word doc). Used 100mL, 8 day old culture inoculated from a plate on [20090505](/Sam%27s+Working+Notebook+Jan-May+2009#sjw20090505). Resuspended pellets in 740uL of TE and took an OD600 via the NanoDrop. Diluted the sample appropriately to an OD600 ~ = 1.0 in a final volume of 740uL TE ([see the last three measurements for OD600 of final dilution](http://eagle.fish.washington.edu/Arabidopsis/20090513%20C.pugetti%20SJW.bmp)).

Followed protocol. Recovered 300uL of aqueous phase prior to precipitation with isopropanol (Step #21). Resuspended DNA in 20uL of H2O. Will run samples on gel according to JGI instructions.

![](https://eagle.fish.washington.edu/Arabidopsis/20090514%20C%20pugetti%20gDNA%20JGI%20QC.jpg)

Lane 1 - 15ng standard (5uL)

Lane 2 - 31ng standard (5uL)

Lane 3 - 63ng standard (5uL)

Lane 4 - Marker 2 (5uL)

Lane 5 - C. pugetti DNA (5uL: 3uL + 2uL dye)

Lane 6 - Marker 3 (5uL)

Lane 7 - 125ng standard

Lane 8 - 250ng standard (5uL)

Lane 9 - 500ng standard (5uL)

Results: DNA looks stellar! Just like the example gel in the JGI QC documentation. however, looks to be too little TOTAL yield of DNA to send for sequencing (need