---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2011-06-03 03:49:50+00:00
layout: post
slug: qpcr-c-gigas-actin-and-gapdh-on-v-vulnificus-exposure-cdna-from-20110311
title: qPCR - C.gigas actin and GAPDH on V.vulnificus exposure cDNA (from 20110311)
categories:
  - 2011
  - Miscellaneous
tags:
  - actin
  - cDNA
  - CFX96
  - Crassostrea gigas
  - GAPDH
  - Pacific oyster
  - qPCR
  - Vibrio vulnificus
---

Ran a qPCR on all cDNA samples from the V.vulnificus exposure experiment from [20110111](/Sam%27s+Working+Notebook+Jan+2011+-+March+2011#sjw20110111). This qPCR was to test 2 of 4 potential normalizing genes to evaluate which genes show the least amount of effect from the treatments in this experiment. Primers for actin used were Cg_Actin_306_F (SR ID: 1170), Cg_Actin_408_R (SR ID: 1171). Samples were run in duplicate. [Master mix calcs are here](https://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20110602-01.jpg). The master mix info is the same that was used earlier today, but with the primers noted above, not those listed on the calcs page. Plate layout, cycling params, etc., can be seen in the qPCR Report (see Results).

Results:

[qPCR Data File](https://eagle.fish.washington.edu/Arabidopsis/qPCR/CFX96/Roberts%20Lab_2011-06-02%2018-24-15_CC009827.pcrd) (BioRad CFX96)

[qPCR Report](https://eagle.fish.washington.edu/Arabidopsis/qPCR/CFX96/Roberts%20Lab_2011-06-02%2018-24-15_CC009827.pdf) (PDF)

Actin: Average Cq = 20.21, Standard Deviation = 1.22

GAPDH: Average Cq = 24.42, Standard Deviation = 0.519

Based on the results from the 4 normalizing genes examined, I will use GAPDH as the normalizing gene due to it having the lowest standard deviation of the 4 normalizing genes. Will perform another qPCR to run a duplicate of GAPDH so that we have a second rep.