---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2011-02-05 04:41:53+00:00
layout: post
slug: qpcr-test-young-lab-qpcr-calibration-2
title: qPCR - Test Young Lab qPCR Calibration
categories:
  - 2011
  - Miscellaneous
tags:
  - 18s
  - ABI 7300
  - Crassostrea gigas
  - gDNA
  - Pacific oyster
  - qPCR
  - 'SR ID: 156'
  - 'SR ID: 157'
  - troubleshooting
  - Young Lab
---

Recently, the Young Lab's ABI 7300 qPCR machine was calibrated. Steven asked me to run a plate and see how well the calibration worked. Ran a plate with C.gigas gDNA and Gigas 18s primers (SR ID: 156 and 157) that are known to amplify gDNA. [Master mix calcs are here (top half of page)(https://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20110204-01.jpg). Cycling params were as follows:




    
  * 95C - 10min



40 Cycles of:


    
  * 95C - 15s

    
  * 55C - 30s

    
  * 72C - 30s



Melt curve.

Results:

Absolutely no amplification of any kind. However, I did use one of our conventional PCR plates and not one of the ABI "prism" plates. Additionally, when I removed the plate from the machine, the plate looked as though it had been vigorously shaken:

![](https://lh6.googleusercontent.com/_Mm7i0Up2xoE/TVAvuZUsOzI/AAAAAAAAHsQ/Woi0u7S68Ik/s400/IMAG0030.jpg)

Will repeat this qPCR with a proper ABI "prism" plate.
