---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2009-02-28 01:12:40+00:00
layout: post
slug: qpcr-new-16s-primers-for-v-tubiashii-control-vs-autoclaved-gigas-samples-see-20090224
title: qPCR - New 16s primers for V.tubiashii Control vs. Autoclaved gigas samples
  (see 20090224)
categories:
  - 2009
  - Miscellaneous
tags:
  - '16s'
  - Opticon2
  - qPCR
  - SensiMix
  - SYBR
  - Vibrio tubiashii
---

qPCR was performed using SensiMix/SYBR "kit" with DNAsed RNA samples from [20090224](/Sam%27s+Working+Notebook+Jan-May+2009#sjw20090224). This qPCR used the new V.tub_16s_V3 primers in hopes of getting better amplification; both in signal intensity and elimination of the double peak seen in the melting curves from [20090224](/Sam%27s+Working+Notebook+Jan-May+2009#sjw20090224). [The plate layou/qPCR workup is here](https://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20090227-1.jpg).

Results: Fluorescence comes up WAY too early; at like the 5th cycle! Also, there are two peaks in the melting curves. Additionally, there is a signal in the two water samples and the melting curve for this contamination matches up with one of the melting cure peaks seen in the actual sample melting curves. So, there is some sort of contamination somehwere. Will repeat this using a clean water for the master mix and hope the problem goes away.