---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2009-06-09 04:44:10+00:00
layout: post
slug: pcr-c-pugetti-gdna-from-20090513-20090526
title: PCR - C.pugetti gDNA from 20090513 & 20090526
categories:
  - 2009
  - Miscellaneous
tags:
  - 1492R
  - 16s
  - 27F
  - Cycloclasticus pugetii
  - gDNA
  - gel
  - PCR
---

Previous PCRs from [20090601](/Sam%27s+Working+Notebook+Jun-Aug+2009#sjw20090601) & [20090602](/Sam%27s+Working+Notebook+Jun-Aug+2009#sjw20090602) both showed contamination in the negative control. Suspect that the primer stocks were contaminated due to the usage of older, non-sterile TE for reconstitution. New stocks were received and reconstituted with filter-sterilized TE. Working stocks were made with filter-sterilized Nanopure H2O. All pipettes, tips, tubes, racks were UV-sterilized in the biological hood. The PCR reaction was set up in the biological hood. [PCR set up is here](https://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20090608-02.jpg). Used universal 16s bacteria primers (27F, 1492R). Sequences from Sara Kelly. Anneal 60C.

![](https://eagle.fish.washington.edu/Arabidopsis/20090609.JPG)

Lane 1 - 100bp ladder

Lane 2 - gDNA (5/13/2009)

Lane 3 - gDNA (5/26/2009)

Lane 4 - H2O

Lane 5 - H2O

Lane 6 - H2O

Lane 7 - H2O

Results: Still contamination in the water-only samples!!!