---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2018-08-09 21:08:36+00:00
layout: post
slug: rna-isolation-quantificaiton-tanner-crab-hemolymph
title: RNA Isolation & Quantificaiton - Tanner Crab Hemolymph
categories:
  - 2018
  - Miscellaneous
tags:
  - Chionoecetes bairdi
  - hemolymph
  - Qubit 3.0
  - Qubit RNA HS
  - RNA
  - RNA isolation
  - RNeasy Plus Micro Kit
  - tanner crab
---

Isolated RNA from [40 Tanner crab hemolymph samples selected by Grace](https://github.com/RobertsLab/resources/issues/336) with the RNeasy Plus Micro Kit (Qiagen) according to [the manufacturer's protocol](https://github.com/RobertsLab/resources/blob/master/protocols/Commercial_Protocols/Qiagen_RNeasy-Plus-Micro-Handbook.pdf), with the following modifications:





  * Added mercaptoethanol (2-ME) to Buffer RLT Plus.



  * All spins were at 21,130g



  * Did _not_ add RNA carrier



  * Used QIAshredder columns to aid in homogenization and removal of insoluble material



  * Eluted with 14uL






RNA was quantified using the Qubit RNA HS (high sensitivity) Assay and run on the Roberts Lab Qubit 3.0.

Used 1uL of sample for quantification.

RNA was returned to the -80C box from where original samples had been stored (Rack 2, Row 3, Column 4).



* * *





##### RESULTS



Qubit quantification (Google Sheet):





  * [20180809_qubit_RNA_crab](https://docs.google.com/spreadsheets/d/1Je45_zXiaZeh478XA5kyFdVQIXiU4rvQeEELfGZTgfY/edit?usp=sharing)



Overall, the results aren't great. Only 15 samples (out of 40) had detectable amounts of RNA. Yields from those 15 samples ranged from 40ng - 300ng, with most landing between 50 - 100ng.

Will pass info along to Grace. Will likely meet with her and Steven to discuss plan on how to move forward.
