---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2011-02-01 04:47:11+00:00
layout: post
slug: genomic-pcr-repeat-of-c-gigas-cox-genomic-pcr-from-20110118
title: Genomic PCR - Repeat of C.gigas COX genomic PCR from 20110118
categories:
  - 2011
  - Miscellaneous
tags:
  - COX
  - Crassostrea gigas
  - cyclooxygenase
  - gel
  - gel extraction
  - Pacific oyster
  - PCR
  - PGS
  - prostaglandin synthase
  - 'SR ID: 1148'
  - 'SR ID: 1149'
  - 'SR ID: 1151'
  - Ultrafree-DA
---

This was repeated to generate more PCR product for sequencing purposes. [PCR master mix calcs and cycling params are here](https://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20110131.jpg). Master mixes 04 and 05 (MM04 and MM05) were repeated to gain more PCR product from the faint 550bp & 1500bp bands(MM04) and 5000bp band (MM05).

MM04 - Cg_COX_982_F (SR ID: 1151) + Cg_COX_1545_R (SR ID: 1148) Band size w/o intron = ~550bp

MM05 - Cg_COX_982_F (SR ID: 1151) + Cg_COX_2138_R (SR ID: 1149) Band size w/o intron = ~1130bp

Results:

Gel was run on 20110203

![](https://eagle.fish.washington.edu/Arabidopsis/20110203%20DNA%20gel.jpg)

Samples on the left portion of the gel are from the MM04 primer combo and those on the right are from the MM05. Boxed bands were excised, purified using Millipore Ultra DA-free spin columns and stored @ -20C in Sam's "Misc. -20C Box."

Interestingly in the MM05 set, inconsistent, faint bands of ~400-500bp are visible. These were not visible the first time this PCR was conducted (see [20110118](https://robertslab.github.io/sams-notebook/posts/2011/2011-01-19-genomic-pcr-c-gigas-cyclooxygenase-cox-genomic-sequence/)), but the exposure of the gel image wasn't turned up as high as in this image. Due to their inconsistency and extremely low yield, these bands were not excised.