---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2015-05-15 06:27:47+00:00
layout: post
slug: qpcr-jakes-o-lurida-ctenidia-dnased-rna-control-samples
title: qPCR – Jake's O.lurida ctenidia DNased RNA (Control Samples)
categories:
  - 2015
  - Miscellaneous
tags:
  - actin
  - ctenidia
  - DNased RNA
  - gill
  - olympia oyster
  - Ostrea lurida
  - qPCR
  - 'SR ID: 1504'
  - 'SR ID: 1505'
  - SsoFast EvaGreen Supermix
---

Ran qPCR on DNased RNA from earlier today to assess whether there was any residual gDNA after [the DNase treatment](https://robertslab.github.io/sams-notebook/posts/2015/2015-05-14-dnase-treatment-jakes-o-lurida-ctenidia-rna-controls-from-20150507/) with Oly_Actin_F/R primers (SR IDs: 1505, 1504).

Used 1μL from all templates.

All samples were run in duplicate.

Positive control was [HL1 _O.lurida_ DNA isolated by Jake on 20150323](https://heareresearch.blogspot.com/2015/03/3-23-2015-ezna-dna-isolation-with-seed.html).

Cycling params:




    
  * 95C – 2.5mins

    
  * 40 cycles of:

    
    * 95C – 10s

    
    * 60C – 20s




    
  * Melt curve



Master mix calcs are here: [20150514_qPCR_Oly_DNased_RNA](https://docs.google.com/spreadsheets/d/1aIPp2edvn4cDHWUWgHC3b5I_1Oef73EP1nSKok0WbN8/edit?usp=sharing)

qPCR Plate Layout: [20150514_qPCR_plate_Jake_Oly_Control_RNA](https://docs.google.com/spreadsheets/d/1o6jFPQe15r7DZ_Z_AVfJIdvf1njM3uhwq8fsvZWu4Uw/edit?usp=sharing)



Results:

qPCR Data File (Opticon): [Sam_20150514_153529.tad](https://eagle.fish.washington.edu/Arabidopsis/qPCR/Opticon/Sam_20150514_153529.tad)

qPCR Report (Google Spreadsheet): [20150514_qPCR_Report_Jake_Oly_DNased_Control_RNA](https://docs.google.com/spreadsheets/d/1f3K75ZRuXDN5W5-nP18Y8OVMqyZVCbmE6j75_PZr1os/edit?usp=sharing)

Positive control comes up around cycle ~21.

No amplification in the no template controls.

Four wells of the DNased RNA samples exhibit amplification (B5, C10, C12, D3), however each respective replicate does not. Will re-test these four samples (NC1, SC1, SC2, SC4).





### Amplification Plots



![](https://eagle.fish.washington.edu/Arabidopsis/20150514_qPCR_Amp_DNased_RNA_Jake_Oly_controls.JPG)(http://eagle.fish.washington.edu/Arabidopsis/20150514_qPCR_Amp_DNased_RNA_Jake_Oly_controls.JPG)





### Melt Curves



![](https://eagle.fish.washington.edu/Arabidopsis/20150514_qPCR_Melt_DNased_RNA_Jake_Oly_controls.JPG)(http://eagle.fish.washington.edu/Arabidopsis/20150514_qPCR_Melt_DNased_RNA_Jake_Oly_controls.JPG)


