---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2018-10-03 20:48:54+00:00
layout: post
slug: rna-isolation-ronits-c-gigas-diploidtriploid-dessicationheat-shock-ctenidia-tissues
title: RNA Isolation - Ronit's C.gigas diploid/triploid dessication/heat shock ctenidia
  tissues
categories:
  - 2018
  - Miscellaneous
tags:
  - Crassostrea gigas
  - ctenidia
  - Pacific oyster
  - RNA isolation
  - RNAzol RT
---

Isolated RNA from a subset of Ronit's _Crassostrea gigas_ ctenidia samples (see [Ronit's notebook for experiment deets](https://genefish.wordpress.com/author/voicesonsocialissues/)):





  * D01 C



  * D02 C



  * D19 C



  * D20 C



  * T01 C



  * T02 C



  * T19 C



  * T20 C






RNA was isolated using RNAzol RT (Molecular Research Center) in the following way:





  * Unweighed, frozen tissue transferred to 500uL of RNAzol RT and immediately homogenized with disposable pestle.



  * Added additional 500uL of RNAzol RT and vortexed to mix.



  * Added 400uL of 0.1% DEPC-treated H2O, vortexed and incubated 15mins at RT.



  * Centrifuged 12,000g for 15mins at RT.



  * Transferred 750uL of supernatant to clean tube (discarded remainder), added 1 volume (750uL) of isopropanol, vortexed, and incubated at RT for 10mins.



  * Centrifuged 12,000g for 10mins at RT.



  * Discarded supernatant.



  * Washed pellet with 75% ethanol (made with 0.1% DEPC-treated H2O).



  * Centrifuged 4,000g for 2mins at RT.



  * Discarded supernatant and repeated wash steps.






Pellet was resuspended in 50uL of 0.1% DEPC-treated H2O and stored @ -80oC in Ronit's temporary box.