---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2011-02-05 04:44:33+00:00
layout: post
slug: qpcr-test-young-lab-qpcr-calibration-repeat-3
title: qPCR - Test Young Lab qPCR Calibration (Repeat)
categories:
  - 2011
  - Miscellaneous
tags:
  - ABI 7300
  - qPCR
  - troubleshooting
  - Young Lab
---

This is a repeat of an earlier run from today, but with a different qPCR plate. [Here're master mix calcs (bottom 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 - 1m



Melt curve.

Results:

All samples amplified and showed a proper dissociation curve. However, it does look like there's a spread of ~3 Cts across the plate. This is not good, as this is the equivalent of ~10-fold difference in gene expression. Will repeat again and see if specific wells show the same relative differences in Cts.
