---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2012-05-01 23:53:55+00:00
layout: post
slug: qpcr-detection-of-v-tubiashii-presence-and-expression-using-vtpa-primers-in-dnacdna-from-yesterday
title: qPCR - Detection of V.tubiashii Presence and Expression Using VtpA Primers
  in DNA/cDNA from yesterday
categories:
  - 2012
  - Miscellaneous
tags:
  - CFX96
  - Crassostrea gigas
  - larvae
  - Pacific oyster
  - qPCR
  - Vibrio tubiashii
  - VtpA
---

Ran qPCR with VtpA primers on cDNA and DNA (from yesterday) of C.gigas larvae to see levels of V.tubiashii compared to their water filter samples (see [20120326](/Sam%27s+Working+Notebook+January+-+March+2012#sjw20120326)). Master mix calcs are here. Plate layout, cycling params, etc can be seen in the qPCR Report (see Results). Used 1uL of cDNA and 100ng (1uL) of DNA as template.

All samples were run in duplicate.

Results:

[qPCR Data File](https://eagle.fish.washington.edu/Arabidopsis/qpcr/CFX96/Roberts%20Lab_2012-05-01%2008-04-41_cc009827.pcrd) (CFX96)
[qPCR Report](https://eagle.fish.washington.edu/Arabidopsis/qpcr/CFX96/Roberts%20Lab_2012-05-01%2008-04-41_CC009827.pdf) (PDF)

No detectable levels of expression (or, no expression at all) in any of the cDNA samples.

Below I've put together a very rough comparison of larvae levels, based off of the the standard curve. I have NOT done the full back calculations!! This is data straight out of the qPCR machine, using the standard curve. Due to the large range, I've graphed the data on a logarithmic scale so all the data is visible on the graph.

![](https://img.skitch.com/20120502-r83eaackyu4iasgxta3uicb7kp.jpg)