---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
layout: post
title: Reverse Transcription - C.gigas PolyIC Diploid MgCl2
date: '2023-08-16 06:37'
tags: 
  - reverse transcription
  - MMLV
  - Crassostrea gigas
  - Pacific oyster
categories: 
  - 2023
  - Miscellaneous
---
After [quantifying diploid MgCl<sub>2</sub>](https://robertslab.github.io/sams-notebook/posts/2023/2023-08-16-RNA-Quantification---C.gigas-PolyIC-Diploid-MgCl2/) (Notebook) earlier today and proceeded to make cDNA. Reverse transcription was performed using oligo dT primers using M-MLV RT (Promega), per the manufacturer's recommendations. Used 400ng of RNA in each reaction. I used 400ng (instead of the usual 100ng) to simplify pipetting for high concentration samples, without the need/time to dilute samples. All reactions were done on ice in 0.5uL PCR tubes.

cDNA was stored in the same -80<sup>o</sup>C box as the original RNA.

- [202300816-cgig-polyIC-cDNA-calcs](https://docs.google.com/spreadsheets/d/15PVvlGGn4xLsmH6uERtCHugk4R9EDMVyMk_VHR109LM/edit?usp=sharing) (Google Sheet)