---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
layout: post
title: RNA Isolation - C.gigas Ctenidia from Noah's Heat-Mechanical Stress Project
date: '2023-07-12 12:27'
tags: 
  - RNA isolation
  - Noah
  - ctenidia
  - Crassostrea gigas
  - Pacific oyster
categories: 
  - 2023
  - Miscellaneous
---
[Steven asked that I help isolate RNA](https://github.com/RobertsLab/resources/issues/1674) (GitHub Issue) for Noah's summer project involving heat and mechanical stress (tumbling) of adult [_Crassostrea gigas_ (Pacific oyster)](http://en.wikipedia.org/wiki/Pacific_oyster). Here's a brief overview of the process:

- Frozen ctenidia tissue was kept on ice.
- Piece of tissue was placed in 500uL RNAzol RT in a 1.7mL tube and immediately homogenized using a disposable pestle.
- An additional 500uL of RNAzol RT was added to the tube, vortexed, and incubated at room temp for ~5mins.
- Insoluble material was pelleted at 21,000g for 1minute.
- Supernatant was transferred to 2mL tube containing 1mL 100% ethanol and vortexed.

Used 700uL of each sample for use in the Direct-zol mini RNA Prep Kit (ZymoResearch). Manufacturer's protocol was followed, including DNase step.

Samples were stored in Noah's -80C project box. RNA will be quantified tomorrow.


| Sample         |
|----------------|
| NTC1_ctenidia  |
| NTC2_ctenidia  |
| NTH1_ctenidia  |
| NTH2_ctenidia  |
| NTHM1_ctenidia |
| NTHM2_ctenidia |
| NTM1_ctenidia  |
| NTM2_ctendia   |
| TC1_ctenidia   |
| TC2_ctendia    |
| TH1_ctenidia   |
| TH2_ctenidia   |
| THM1_ctenidia  |
| THM2_ctenidia  |
| TM1_ctendia    |
| TM2_ctenidia   |