---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2009-12-16 00:16:21+00:00
layout: post
slug: reverse-transcription-abalone-0712-dnased-rna-from-20090623
title: Reverse Transcription - Abalone 07:12 DNased RNA (from 20090623)
categories:
  - 2009
  - Miscellaneous
tags:
  - '07:12'
  - DNased RNA
  - M-MLV
  - NanoDrop1000
  - oligo dT
  - reverse transcription
  - RNA quantification
---

Spec - DNased Abalone 07:12 RNA

Apparently, these samples had not been spec'd after DNase treatment.

Results:

![](https://eagle.fish.washington.edu/Arabidopsis/RNA%20Spec%20Readings/20091215%20DNased%20RNA%20SJW.jpg)

Samples range in quality (260/280) from not great to perfect. Will perform calcs to make cDNA.



Set up reverse transcription rxns using 174ng of each DNased RNA (sample 07:12-04 was limiting; only 6.1uL available), using Promega oligo dT primers and M-MLV Reverse Transcriptase according to Promega protocol. [RNA/oligo dT primer workup is here](mailto:Primer%20and%20DNAsed%20RNA%20were%20mixed%20and%20brought%20up%20to%2018.25uL%20with%20H2O.%20Samples%20were%20heated%20@%2070C%20for%205mins%20and%20then%20placed%20immediately%20on%20ice.). Primer and DNAsed RNA were mixed and brought up to 18.25uL with H2O. Samples were heated @ 70C for 5mins and then placed immediately on ice. The RT Master Mix set up can be found here. 6.75uL of the RT master mix was added to each tube, mixed, spot spun, incubated 42C for 1hr., 95C for 3mins and then the samples were given to Lisa in the Friedman Lab.
