---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2011-08-26 02:45:27+00:00
layout: post
slug: pcr-full-length-pgs1-pgs2-cdnas
title: PCR - Full-length PGS1 & PGS2 cDNAs
categories:
  - 2011
  - Miscellaneous
tags:
  - cDNA
  - Crassostrea gigas
  - gel
  - gel extraction
  - Pacific oyster
  - PCR
  - PGS
  - PGS1
  - PGS2
  - Ultrafree-DA
---

Ran PCR to amplify full-length cDNAs of PGS1 & PGS2 (COX1 & COX2) using primers designed to anneal in the 5'/3'UTRs of each isoform. PGS1 primers = SRIDs: 1377, 1378. PGS2 primers = 1376, 1375. [Master mix calcs and cycling params are here](https://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20110825-01.jpg). cDNA was pooled cDNA made [20110311](/Sam%27s+Working+Notebook+Jan+2011+-+March+2011#sjw20110311) from various tissues.

PGS1 Expected Size = ~2300bp

PGS2 Expected Size = ~2500bp

Results:

![](https://eagle.fish.washington.edu/Arabidopsis/20110825-01.jpg)

Gel

Lane 1 - Hyperladder I (Bioline)

Lane 2 - PGS1

Lane 3 - PGS1 NTC

Lane 4 - PGS1 NTC

Lane 5 - PGS2

Lane 6 - PGS2 NTC

Lane 7 - PGS2 NTC

PGS1 Results: PGS1 PCR produces a single band of the expected size (~2300bp), indicating that the two primers, which were designed to anneal in the 5'/3'UTRs of the gene and should be highly specific to just this isoform, work perfectly. The band was excised and stored @ -20C in "Sam's Miscellaneous" box.

PGS2 Results: PGS2 PCR didn't produce any product. Will repeat with a lower annealing temp (50C instead of 55C).
