---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2015-05-15 06:28:09+00:00
layout: post
slug: qpcr-jakes-o-lurida-ctenidia-dnased-rna-1hr-heat-shock-samples
title: qPCR – Jake's O.lurida ctenidia DNased RNA (1hr Heat Shock Samples)
categories:
  - 2015
  - Miscellaneous
tags:
  - actin
  - ctenidia
  - DNased RNA
  - gill
  - olympia oyster
  - Opticon
  - Opticon2
  - 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/2015/2015-05-15-dnase-treatment-jakes-o-lurida-ctenidia-rna-1hr-heat-shock-from-20150506/) 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 (used same calcs from the other day): [20150512_qPCR_Oly_RNA](https://docs.google.com/spreadsheets/d/1-jUGGyD56GcA_uk07TFUEh2R0Y2e6DxeEzqdByTccJE/edit?usp=sharing)

Plate layout: [20150514_qPCR_plate_Jake_Oly_1hr_HS_DNased_RNA](https://docs.google.com/spreadsheets/d/1A4x_0wd2EhMo84msvb-dsLluARUU8ily0h6IQL_5V_k/edit?usp=sharing)



Results:

qPCR Data File (Opticon): [Sam_20150514_170332.tad](https://eagle.fish.washington.edu/Arabidopsis/qPCR/Opticon/Sam_20150514_170332.tad)

qPCR Report (Google Spreadsheeet): [20150514_qPCR_Report_Jake_Oly_DNased_1hr_HS_RNA](https://docs.google.com/spreadsheets/d/1DY3eA1Wh01TrcZ8DFX5Ij-EEDhPimYmWfUvsYGaxEc4/edit?usp=sharing)



Positive control samples are the only samples that produced amplification (cycle ~20). Will proceed to making cDNA.





### Amplification Plots



![](https://eagle.fish.washington.edu/Arabidopsis/20150514_qPCR_Amp_DNased_RNA_Jake_Oly_1hr_HS.JPG)(http://eagle.fish.washington.edu/Arabidopsis/20150514_qPCR_Amp_DNased_RNA_Jake_Oly_1hr_HS.JPG)





### Melt Curves



![](https://eagle.fish.washington.edu/Arabidopsis/20150514_qPCR_Melt_DNased_RNA_Jake_Oly_1hr_HS.JPG)(http://eagle.fish.washington.edu/Arabidopsis/20150514_qPCR_Melt_DNased_RNA_Jake_Oly_1hr_HS.JPG)
