---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2009-12-23 23:57:02+00:00
layout: post
slug: reverse-transcription-bb-dh-dnased-rna-from-20090514
title: Reverse Transcription - BB & DH DNased RNA (from 20090514)
categories:
  - 2009
  - PROPS
tags:
  - BB
  - cDNA
  - Crassostrea gigas
  - DH
  - DNased RNA
  - M-MLV
  - oligo dT
  - Pacific oyster
  - reverse transcription
---

Made a fresh, double batch (50uL rxn instead of 25uL) of cDNA according to Promega MMLV protocol using oligo dT primers. cDNA was put into a plate for faster qPCR loading. [cDNA calcs and plate layout are here](https://spreadsheets.google.com/ccc?key=0AmS_90rPaQMzdFVBS3FUQy12a1ZMekZ4OC1vTnA2aGc&hl=en). Briefly, RNA and oligo dTs were combined, brought up to 37uL, heated @ 70C for 5mins and immediately placed on ice. RT master mix was made ([RT master mix calcs are here](http://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20091223-01.jpg)), 13uL was distributed to each well. Samples were incubated @ 42C for 1hr and then 95C for 5mins.

UPDATE: cDNA plate was discarded 20120320 by SJW.