---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2011-07-27 03:04:22+00:00
layout: post
slug: colony-pcrs-c-gigas-cox2pgs2-clones-from-yesterday
title: Colony PCRs - C.gigas COX2/PGS2 Clones (from yesterday)
categories:
  - 2011
  - Miscellaneous
tags:
  - colony screen
  - COX
  - COX2
  - Crassostrea gigas
  - cyclooxygenase
  - gel
  - Pacific oyster
  - PCR
  - PGS
  - PGS2
  - prostaglandin synthase
---

Performed colony PCRs on the 4 sets of cloning reactions that were performed yesterday using the M13F/R vector primers. Colonies were picked, restreaked on a fresh LB Kan50 plates (made 20110726 by SJW) and PCR'd. [Master mix calcs are here](https://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20110726-01.jpg). Selected 8 white colonies from each cloning reaction for PCR. Restreaked plate was incubated @ 37C O/N.

Cycling Params:




    
  * 95C - 10m



40cycles of:


    
  * 95C - 10s

    
  * 55C - 10s

    
  * 72C - 3m



Results:

![](https://eagle.fish.washington.edu/Arabidopsis/20110727-01%20Gel.jpg)

![](https://eagle.fish.washington.edu/Arabidopsis/20110727-02%20Gel.jpg)

Hyperladder I is used as the ladder in both gels.

Cloning results look great (except Colony #1 in the 5' Top Band didn't produce a product). Will select a re-streaked colony from each set and inoculate liquid culture for mini prep and subsequent sequencing.
