---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
layout: post
title: DNA Isolation - C.gigas Ploidy Experiment Ctenidia
date: '2018-12-20 08:38'
tags:
  - Crassostrea gigas
  - Pacific oyster
  - DNA isolation
  - E.Z.N.A. Mollusc DNA Kit
  - Qubit 3.0
  - Qubit dsDNA BR Assay
categories:
  - 2018
  - Miscellaneous
---
Yesterday, Ronit initiated DNA isolation from ctenidia samples [from his experiment (Google Sheet)](https://docs.google.com/spreadsheets/d/17mv8gMbmaldggA8Zf0RwBeNF_O4faY8dJFg31XO63K4/edit?usp=sharing) from the following four samples:

- D11
- D12
- D13
- D14

Frozen tissue was excised from frozen tissue block via razor blade (weight not recorded) and pulverized under liquid nitrogen. Samples were incubated O/N @ 37<sup>o</sup>C (heating block) in 350uL of MB1 Buffer + 25uL Proteinase K, per the E.Z.N.A. Mollusc DNA Kit (Omega) instructions.

After the O/N incubation, I processed the samples according to the E.Z.N.A. Mollusc DNA Kit (Omega) with the following notes:



Samples were eluted in 100uL of Elution Buffer and quantified with the Roberts Lab Qubit 3.0 and the Qubit dsDNA BR Assay (Invitrogen), using 2uL of each sample.

Samples were stored in "[Ronit's gDNA Box #1 (positions B2 - B5)](https://docs.google.com/spreadsheets/d/1Gp91BJ5g2W6c4r8UhGVc2UhQGdCHju72aZswpR-Qxpc/edit?usp=sharing)"  in the FTR213 -20<sup>o</sup>C freezer.

---

# RESULTS

Qubit data (Google Sheet):

- [20181220_qubit_gDNA_gigas_ploidy](https://docs.google.com/spreadsheets/d/1KFkA9hL217watfnJoC3DNq9R9OPm7n_Az3vRG8WLWdY/edit?usp=sharing)



| sample_ID | Concentration(ng/uL) |
|-----------|----------------------|
| D11-C     | 181                  |
| D12-C     | 192                  |
| D13-C     | 288                  |
| D14-C     | 452                  |