---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2011-03-04 23:29:18+00:00
layout: post
slug: 3race-c-gigas-3race-for-cox2
title: 3'RACE - C.gigas 3'RACE for COX2
categories:
  - 2011
  - Miscellaneous
tags:
  - cDNA
  - COX
  - COX2
  - Crassostrea gigas
  - cyclooxygenase
  - gel
  - Pacific oyster
  - PGS
  - PGS2
  - prostaglandin synthase
  - RACE
  - RACE PCR
  - SMART RACE
---

Used Cg_COX2_3'RACE_short (SR ID: 1197) & Cg_COX2_3'RACE_long (SR ID: 1196) and the Clonetech SMART RACE cDNA Amplification Kit (unknown acquisition date) to attempt to acquire more 3' sequence of the C.gigas COX2 isoform. Used Gigas 3'RACE cDNA (from 20080610).

Results:

![](https://eagle.fish.washington.edu/Arabidopsis/20110307-01.jpg)

Gel Loading:

Lane 1: Hyperladder 1

Lane 2: empty

Lane 3: Cg_COX2_3'RACE_long

Lane 4: Cg_COX2_3'RACE_long NTC

Lane 5: empty

Lane 6: Cg_COX2_3'RACE_short

Lane 7: Cg_COX2_3'RACE_short NTC

Lane 8: Hyperladder 1

No products produced. This could be due to a large number of factors. The age of the cDNA (from 20080610) is well beyond what the Clontech manual says for storage term (6 months). Additionally, the Clontech polymerase used was nearly 6 years old. The kit (and its components) are of an unknown age and could factor in to the failure of this procedure. Also, the primers that were designed had less than ideal Tm, per the kit's recommendations.

May need to sequence some previously purified potential COX2 fragments in order to obtain a more useable region of the gene for RACE.
