---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
layout: post
title: DNase - Ronit's C.gigas Ctenidia RNA
date: '2018-11-15'
tags:
  - ctenidia
  - Crassostrea gigas
  - Pacific oyster
  - RNA
  - DNase
  - Turbo DNA-free
  - diploid
  - triploid
categories:
  - 2018
  - Miscellaneous
---
Ronit finished his RNA isolations for his ctenidia samples on [20181025](https://genefish.wordpress.com/2018/10/25/ronits-notebook-rna-extraction-for-c-gigas-desiccation-elevated-temperature-samples-round-2/) & [20181101](https://genefish.wordpress.com/2018/10/25/ronits-notebook-rna-extraction-for-c-gigas-desiccation-elevated-temperature-samples-round-2/), as well as quntification (he has no notebook entry for this, at this time). However, his Qubit data can be found in these two Google Sheets:

- [20181108_qubit_RNA_ronit_ctenidia](https://docs.google.com/spreadsheets/d/1fb3nVN85sMEAO5wM8O6li8Z9vDQXVXIAenzmig2WHUc/edit?usp=sharing)

- [20181114_qubit_RNA_ronit_ctenidia_1-10_dilution](https://docs.google.com/spreadsheets/d/1guXLs2mq0uV8bXmEET2HAlxrT_isBa5UL1Y3yy0KgR8/edit?usp=sharing)

In preparation for reverse transcription, I DNased his samples using the Turbo DNA-free Kit (Ambion) according to the manufacturer's standard protocol.

Used 500ng of each sample.

NOTE: Sample T06 was very dilute, so performed a 60uL reaction instead of the standard 50uL reaction.

Calculations are here (Google Sheet):

- [20181115_DNase_ronit_RNA_calcs](https://docs.google.com/spreadsheets/d/13GLxHpUqUpeHkAG_mozxHRveNY61DDz5Tau2HcniRgY/edit?usp=sharing)

Ronit's master sheet (Google Sheet) is here:

- [Exposure 8/29-8/30 C.Gigas](https://docs.google.com/spreadsheets/d/17mv8gMbmaldggA8Zf0RwBeNF_O4faY8dJFg31XO63K4/edit?usp=sharing)

Will check these via qPCR to verify whether or not they have any detectable gDNA.
