---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2012-01-26 01:56:07+00:00
layout: post
slug: qpcr-dnased-rna-from-earlier-today
title: qPCR - DNased RNA from earlier today
categories:
  - 2012
  - Miscellaneous
tags:
  - 18s
  - CFX96
  - Crassostrea gigas
  - DNased RNA
  - gDNA
  - Pacific oyster
  - qPCR
---

Checked DNased RNA samples from earlier today for the presence of residual gDNA. Used C.gigas BB16 gDNA (from [20110201](/Sam%27s+Working+Notebook+Jan+2011+-+March+2011)) diluted to ~7ng/uL as a positive control to match the dilution factor of the RNA that will be used in the reverse transcription reaction (175ng in 25uL = 7ng/uL). All samples were run in duplicate. [Master mix calcs are here](https://eagle.fish.washington.edu/Arabidopsis//Notebook%20Workup%20Files/20120125-01.jpg). Plate layout, cycling params, etc can be found in the qPCR Report (see Results).

Results:

[qPCR Data](https://eagle.fish.washington.edu/Arabidopsis/qPCR/CFX96/Roberts%20Lab_2012-01-25%2014-52-29_CC009827.pcrd) (CFX96)

[qPCR Report](https://eagle.fish.washington.edu/Arabidopsis/qPCR/CFX96/Roberts%20Lab_2012-01-25%2014-52-29_CC009827.pdf) (PDF)

Positive control (in Green in qPCR Report) worked perfectly and showed excellent repeatability. The remainder of the samples (in Blue in qPCR Report) and the NTCs (in Red in qPCR Report) were extremely inconsistent with many having one replicate show late amplification, while the other replicate showed no amplification at all. Will have to repeat to get a more definitive assessment of residual gDNA content in these samples.
