---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2009-01-23 03:10:38+00:00
layout: post
slug: mrna-isolation-hard-clam-gill-and-hemo-rna
title: mRNA Isolation - Hard Clam gill and hemo RNA
categories:
  - 2009
  - Miscellaneous
tags:
  - gill
  - Hard clam
  - hemocyte
  - Mercenaria mercenaria
  - Micro PolyA Purist
  - mRNA
  - mRNA enrichment
  - NanoDrop1000
  - RNA quantification
---

mRNA was isolated according to [Ambion PolyA Purist protocol](https://aquacul4.fish.washington.edu/Protocols:Information%20Sheets/Commercial%20Protocols:Manuals/Ambion%20-%20MicroPoly%28A%29Purist%20Kit.pdf). After mixing samples with resin, samples were incubated @ RT for 1hr. Samples were washed per the protocol. However, the hemo sample was not clearing from the spin columns with the protocol-directed 3 min. spins. The column had to be spun up to 15 mins. in order for the column to clear. :(

![](https://eagle.fish.washington.edu/Arabidopsis/RNA%20Spec%20Readings/20090122%20RNA%20SJW-02.png)

Results: The gill mRNA looks good (~1.8ug). The concentration of the hemo sample is extremely low and is below the error threshold for the NanoDrop, but it may not be that bad. The samples are in a relatively large volume (~200uL) and the yield is expected to be very small. So, I will precipitate these samples O/N @ -20C according to [Ambion PolyA Purist protocol](https://aquacul4.fish.washington.edu/Protocols:Information%20Sheets/Commercial%20Protocols:Manuals/Ambion%20-%20MicroPoly%28A%29Purist%20Kit.pdf) in order to concentrate them.
