---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2015-05-15 00:46:57+00:00
layout: post
slug: dnase-treatment-jakes-o-lurida-ctenidia-rna-1hr-heat-shock-from-20150506
title: DNase Treatment - Jake's O.lurida Ctenidia RNA (1hr Heat Shock) from 20150506
categories:
  - 2015
  - Miscellaneous
tags:
  - DNase
  - DNased RNA
  - NanoDrop1000
  - olympia oyster
  - Ostrea lurida
  - RNA
  - RNA quantification
  - Turbo DNA-free
---

Since the [_O.lurida_ RNA I isolated on 20150506](https://robertslab.github.io/sams-notebook/posts/2015/2015-05-06-rna-isolation-jakes-o-lurida-ctenidia-1hr-heat-stress-from-20150422/) showed [residual gDNA via qPCR](https://robertslab.github.io/sams-notebook/2015/05/12/qpcr-jake-o-lurida-ctenidia-rna-heat-shock-samples-from-20150506/), I treated 1.5μg of RNA from each sample using the Turbo DNA-free Kit (Ambion/Life Technologies), following the "rigorous" protocol.

Briefly:




    
  * 50μL reactions were carried out in 0.5mL tubes

    
  * added 1μL of DNase to each tube

    
  * incubated 30mins @ 37C

    
  * added additional 1μL of DNased

    
  * incubated 30mins @ 37C

    
  * added 0.2 vols (10.2μL) of DNase Inactivation Reagent

    
  * incubated and mixed for 2mins @ RT

    
  * transferred 50μL of supe to sterile 1.5mL snap cap tubes

    
  * spec'd on Roberts Lab NanoDrop1000



Samples were stored @ -80C in [Shellfish RNA Box #5 and Box #6](https://docs.google.com/spreadsheet/ccc?key=0AmS_90rPaQMzcHdyU1d0MDVMLWpaTWdadnJSd0M4UUE&usp=sharing).

DNase reaction calcs: [20150514_Jake_Oly_1hr_HS_DNase_calcs](https://docs.google.com/spreadsheets/d/1KS3tJand0vKSs6ZJk9t-hChZYmM0--RhcXiR8gDOlYo/edit?usp=sharing)







Results:

Google Spreadsheet: [20150514_DNased_RNA_Jake_Oly_1hr_HS_ODs](https://docs.google.com/spreadsheets/d/1qzfmdoxPG6nP3F5jB23QSREy1S72FxnBW8jwSdxqjZE/edit?usp=sharing)



![](https://eagle.fish.washington.edu/Arabidopsis/20150514_DNased_RNA_Jake_oly_1hr_HS_ODs.JPG)(http://eagle.fish.washington.edu/Arabidopsis/20150514_DNased_RNA_Jake_oly_1hr_HS_ODs.JPG)



![](https://eagle.fish.washington.edu/Arabidopsis/20150514_DNased_RNA_Jake_oly_1hr_HS_plots_01.JPG)(http://eagle.fish.washington.edu/Arabidopsis/20150514_DNased_RNA_Jake_oly_1hr_HS_plots_01.JPG)



![](https://eagle.fish.washington.edu/Arabidopsis/20150514_DNased_RNA_Jake_oly_1hr_HS_plots_02.JPG)(http://eagle.fish.washington.edu/Arabidopsis/20150514_DNased_RNA_Jake_oly_1hr_HS_plots_02.JPG)



All samples look pretty good except for HT1 8 (RNA concentration is ridiculously high!) and NT1 8 (RNA concentration is way below expected). Will check for residual gDNA via qPCR.
