---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2011-10-07 02:20:35+00:00
layout: post
slug: pcr-region-outside-of-coxpgs-qpcr-primers
title: PCR - Region Outside of COX/PGS qPCR Primers
categories:
  - 2011
  - Miscellaneous
tags:
  - COX
  - COX1
  - COX2
  - Crassostrea gigas
  - cyclooxygenase
  - gel
  - gel extraction
  - Pacific oyster
  - PCR
  - PGS
  - PGS1
  - PGS2
  - prostaglandin synthase
  - Ultrafree-DA
---

Ran PCR using primers Cg_COX_982_F and Cg_COX_2138_R (SR IDs: 1149 & 1151, respectively). Template was pooled cDNA from 20110311 of various C.gigas tissues. These primers anneal 5' and 3' of where the qPCR primers for both COX1/PGS1 and COX2/PGS2 anneal. [Master mix calcs and cycling params are here](https://eagle.fish.washington.edu/Arabidopsis/20111006-02.jpg). Ran multiples of the same reaction to ensure sufficient product for use in cloning/PCR.

Results:

![](https://eagle.fish.washington.edu/Arabidopsis/20111006-01.JPG)

Gel is loaded with Hyperladder I (Bioline) and 7 samples (no NTC; don't ask). Band in each lane is of the expected size (~1200bp). Each band was excised and purified using Ultra-free DA columns (Millipore), according to protocol. Purified DNA will be used in a subsequent PCR using the qPCR primers for COX/PGS 1&2 BEFORE cloning this product for sequencing.
