---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2018-10-16 17:33:47+00:00
layout: post
slug: dnase-treatment-ronits-c-gigas-ploiyddessication-ctenidia-rna
title: DNase Treatment - Ronit's C.gigas Ploiyd/Dessication Ctenidia RNA
categories:
  - 2018
  - Miscellaneous
tags:
  - Crassostrea gigas
  - ctenidia
  - dessication
  - diploid
  - DNase
  - Pacific oyster
  - RNA
  - triploid
  - Turbo DNA-free
---

After [quantifying Ronit's RNA earlier today](https://robertslab.github.io/sams-notebook/posts/2018/2018-10-16-rna-quantification-ronits-c-gigas-ploidydessication-rna/), I DNased them using the Turbo DNA-free Kit (Ambion), according to the [manufacturer's standard protocol](https://github.com/RobertsLab/resources/blob/master/protocols/Commercial_Protocols/Ambion_Turbo_DNA_Free.pdf).

Used 1000ng of RNA in a 50uL reaction in a 0.5mL thin-walled snap cap tube. Samples were mixed by finger flicking and then incubated 30mins @ 37oC in a PTC-200 thermal cylcer (MJ Research), without a heated lid.

DNase inactivation was performed (0.1 volumes of inactivation reagent; 5uL), pelleted, and supe transferred to new 1.7mL snap cap tube.

Samples were stored on ice in preparation for qPCR to test for residual gDNA.

DNase calculations are here:





  * [20181016_DNase_calcs](https://docs.google.com/spreadsheets/d/1rgS3MH5QjFYIQL8PpvUf6xo-ZXI8KTX4pm16cb7if4U/edit?usp=sharing)



Samples will be permanently stored here (Google Sheet):



  * [Ronit's Ploidy/Dessication RNA Box #1 - Rack 15, Column 4, Row 4 in -80oC](https://docs.google.com/spreadsheets/d/1Qsvz3QTURlPF_hX05BQxjom3484WuMfqQ1ILl9LEljU/edit?usp=sharing)