---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
layout: post
title: RNA Isolation and Quantification - C.bairdi Hemocyte Pellets in RNAlater
date: '2020-01-26 10:01'
tags:
  - Chionoecetes bairdi
  - Quick DNA-RNA Microprep Plus
  - Tanner crab
  - Qubit3.0
  - hsRNA assay
categories:
  - 2020
  - Tanner Crab RNAseq
---
Isolated RNA from the following  hemolymph pellet samples:

- 6202_337_12

- 6142_474_26

- 6144_477_26

- 6122_489_26

- 6176_490_26

- 6136_491_26

- 6137_500_26

- 6125_503_26

- 6145_505_26

- 6131_507_26

- 6159_508_26

- 6206_509_26

- 6150_511_26

- 6161_512_26

- 6115_514_26

- 6179_515_26

- 6172_517_26

Isolated RNA using the [Quick DNA/RNA Microprep Kit](https://github.com/RobertsLab/resources/blob/master/protocols/Commercial_Protocols/ZymoResearch_quick-dna-rna_microprep_plus_kit_20190411.pdf) (ZymoResearch; PDF) according to the manufacturer's protocol for liquids/cells in RNAlater.

- Used 35uL from each RNAlater/hemocyte slurry.

- Mixed with equal volume of H<sub>2</sub>O (35uL).

- Retained DNA on the Zymo-Spin IC-XM columns for isolation after RNA isolation.

- Performed on-column DNase step.

- RNA was eluted in 15uL H<sub>2</sub>O

RNA was quantified on the Roberts Lab Qubit 3.0 using the RNA High Sensitivity Assay (Invitrogen), using 2uL of each sample.

RNA was stored in the [-80<sub>o</sup>C freezer](http://b.link/srlab-80C) in [Shellfish Box #8](https://docs.google.com/spreadsheets/d/1ax6C-muxUTXxFEtfWdswBvueLhmxZzmwZcO2ur-0q-Q/edit#gid=1794729843).

---

# RESULTS

Qubit restuls (Google Sheet):

- [20200126_qubit_cbai_RNA](https://docs.google.com/spreadsheets/d/1KTqkx4s1eZG2Y0gtqHvW9J9h9ebRBTYZp35NzlQCsMs/edit?usp=sharing)


| Sample ID   | [RNA](ng/uL) |
|-------------|--------------|
| 6202_337_12 | 23.7         |
| 6142_474_26 | 14.4         |
| 6144_477_26 | 20.3         |
| 6122_489_26 | 8.32         |
| 6176_490_26 | 10.7         |
| 6136_491_26 | 7.37         |
| 6137_500_26 | 13.8         |
| 6125_503_26 | 7.12         |
| 6145_505_26 | 4.7          |
| 6131_507_26 | 8.9          |
| 6159_508_26 | 7.74         |
| 6206_509_26 | 9.75         |
| 6150_511_26 | 4.12         |
| 6161_512_26 | 10.9         |
| 6115_514_26 | 5.96         |
| 6179_515_26 | 7.23         |
| 6172_517_26 | Out of range |