---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2009-12-10 00:29:08+00:00
layout: post
slug: reverse-transcription-herring-liver-mrna-for-solid-libraries
title: Reverse Transcription - Herring Liver mRNA for SOLiD Libraries
categories:
  - 2009
  - Miscellaneous
tags:
  - 2L
  - 3L
  - 4L
  - 6L
  - cDNA
  - gel
  - library prep
  - liver
  - MiniElute PCR Purification Kit
  - Pacific herring
  - reverse transcription
  - SOLiD
  - SOLiD libraries
  - Whole Transcriptome Analysis Kit
---

Ligation reactions from yesterday were subject to reverse transcription according to protocol. [Master mix workup info is here](https://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20091209-01.jpg). Samples were incubated @ 42C, 30mins and then cleaned up using the Qiagen MiniElute PCR Purification Kit, according to the Ambion WTK protocol.

After RT rxn, samples were run on a Novex 6% TBE-Urea gel according to Ambion WTK protocol. Samples were loaded, left to right: 2L, 3L, 4L, 6L. Ladders are to the left of each sample. The break in the smear in the 6L sample is a tear in the gel.

![](https://eagle.fish.washington.edu/Arabidopsis/20091209-01.jpg)

The recommended range of cDNA (100-200bp) were excised from the gel and were cut into four pieces, according to the Ambion WTK protocol. The two outer gel slices from each sample will be stored @-20C. Then proceeded to the emulsion PCR. Here is an image of the gel after the cDNA was cut out:

![](https://eagle.fish.washington.edu/Arabidopsis/20091209-02.jpg)