---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2009-04-08 23:16:51+00:00
layout: post
slug: cdna-abalone-rna-from-20090331-20090402
title: cDNA - Abalone RNA from 20090331 & 20090402
categories:
  - 2009
  - Miscellaneous
tags:
  - '08:3'
  - '08:4'
  - black abalone
  - cDNA
  - Haliotis cracherodii
  - Quantitect Kit
  - reverse transcription
  - RNA
---

cDNA was made from the above RNA samples using the Qiagen Quantitect RT Kit. The samples were [laid out in a PCR plate](https://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20090408-01.jpg). 274.2ng of RNA was used in the rxn for each sample, based on the lowest concentration RNA sample (08:3-20) to equalize all the samples. The Genomic Wipeout step of the kit requires 2uL of Genomic Wipeout enzyme/buffer to be added to 12uL of an RNA sample, so [the calculations were done and can be found here](http://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20090408-02.jpg). A check mark on the calculation sheet indicates that the water and then the RNA was added to the appropriate wells. Those with two check marks were used for a "No RT" rxn and thus, have duplicate wells ([see plate layout](http://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20090408-01.jpg)).

The plate was mixed, spot spun, uncubated at 42C for 2mins and immediately placed on ice.

The [RT and No RT master mixes](https://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20090408-04.jpg) were set up on ice and then added to the respective wells ([see sheet here](http://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20090408-03.jpg)).

UPDATE: cDNA plate was discarded 20120320 by SJW.