---
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: '2020-01-24 14:59'
tags:
  - Tanner crab
  - RNA isolation
  - RNA quantification
  - Qubit3.0
  - Qubit hsRNA assay
  - Chionoecetes bairdi
  - Quick DNA-RNA Microprep Plus
categories:
  - 2020
  - Tanner Crab RNAseq
---
Isolated RNA from the following  hemolymph pellet samples:

- 6174_233_12

- 6258_261_12

- 6270_269_12

- 6161_302_12

- 6115_307_12

- 6206_319_12

- 6241_323_12

- 6158_324_12

- 6272_338_12

- 6152_344_12

- 6275_372_12

- 6218_373_12

- 6153_404_26

- 6132_406_26

- 6162_407_26

- 6178_410_26

- 6163_424_26

- 6140_429_26

- 6174_433_26

- 6199_445_26

- 6156_446_26

- 6106_450_26

- 6151_454_26

- 6177_456_26

- 6119_461_26

- 6124_464_26

- 6134_465_26

- 6168_466_26

- 6152_467_26

- 6121_472_26

- 6205_475_26

- 6155_476_26

- 6143_479_26

- 6158_483_26

- 6169_486_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 RNA Box #7](https://docs.google.com/spreadsheets/d/1ax6C-muxUTXxFEtfWdswBvueLhmxZzmwZcO2ur-0q-Q/edit#gid=1549162576) and [Shellfish Box #8](https://docs.google.com/spreadsheets/d/1ax6C-muxUTXxFEtfWdswBvueLhmxZzmwZcO2ur-0q-Q/edit#gid=1794729843).

---

# RESULTS

Qubit restuls (Google Sheet):

- [20200124_qubit_cbai_RNA](https://docs.google.com/spreadsheets/d/1XKruSvdXM66L9UXGS4oPsu5xpppToVDW9zR3NtSsc8s/edit?usp=sharing)


| Sample ID   | [RNA](ng/uL) |
|-------------|--------------|
| 6258_261_12 | 10.6         |
| 6270_269_12 | 31.7         |
| 6161_302_12 | 8.04         |
| 6115_307_12 | 7.89         |
| 6206_319_12 | 11.2         |
| 6241_323_12 | 22.2         |
| 6158_324_12 | 16.5         |
| 6272_338_12 | 10.6         |
| 6152_344_12 | 6.65         |
| 6275_372_12 | 10.1         |
| 6218_373_12 | 14.3         |
| 6174_233_12 | 17.8         |
| 6153_404_26 | 12.2         |
| 6132_406_26 | 12.3         |
| 6162_407_26 | 14.5         |
| 6178_410_26 | Out of range |
| 6163_424_26 | 21.4         |
| 6140_429_26 | 12           |
| 6174_433_26 | 17.6         |
| 6199_445_26 | 20.4         |
| 6156_446_26 | 20.7         |
| 6106_450_26 | 25.4         |
| 6151_454_26 | 11.4         |
| 6177_456_26 | 19.9         |
| 6119_461_26 | 11.2         |
| 6124_464_26 | 19.1         |
| 6134_465_26 | 9.1          |
| 6168_466_26 | 18.4         |
| 6152_467_26 | 5.45         |
| 6121_472_26 | 8.57         |
| 6205_475_26 | 13.1         |
| 6155_476_26 | 15.3         |
| 6143_479_26 | 15.3         |
| 6158_483_26 | 13.3         |
| 6169_486_26 | 5.87         |
