---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
layout: post
title: DNA Isolation and Quantification - Ronit's C.gigas Ploidy Ctenidia
date: '2019-01-23 09:48'
tags:
  - DNA isolation
  - E.Z.N.A Mollusc DNA Kit
  - ctenidia
  - DNA quantification
  - qubit
  - Qubit3.0
  - Qubit dsDNA BR Kit
  - Crassostrea gigas
  - Pacific oyster
  - ploidy
  - diploid
  - triploid
categories:
  - 2019
  - Miscellaneous
---
Last week, I [isolated DNA from all of Ronti's ctenidia samples](https://robertslab.github.io/sams-notebook/posts/2019/2019-01-17-DNA-Isolation---C.gigas-Ploidy-Experiment-Ctenidia/), however one sample (D18-C) didn't isolate properly. So, I performed another isolation procedure with another section of frozen tissue. Tissue was excised from frozen tissue block via razor blade (weight not recorded) and pulverized under liquid nitrogen. Samples were incubated O/N @ 37oC (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:

Sample was eluted in 150uL of Elution Buffer.

Sample was stored in [Ronit's gDNA Box #1 (position B9)](https://docs.google.com/spreadsheets/d/1Gp91BJ5g2W6c4r8UhGVc2UhQGdCHju72aZswpR-Qxpc/edit?usp=sharing) (Google Sheet).

I then thawed all the other ctenidia samples isolated last week and quantified them using the Roberts Lab Qubit3.0, using the Qubit dsDNA BR Kit (Invitrogen). I used 1uL of each sample.

---

# RESULTS

Qubit data (Google Sheet):

- [20190123_qubit_DNA_gigas_ploidy](https://docs.google.com/spreadsheets/d/11iFZsMtK8J8z1zgXChNt9WS0hVrQCPBmVUzP4ti5oXA/edit?usp=sharing)

DNA is present in all samples, ranging from 6.48ng/uL to 388ng/uL.

All data was entered in [Ronit's master sheet](https://docs.google.com/spreadsheets/d/17mv8gMbmaldggA8Zf0RwBeNF_O4faY8dJFg31XO63K4/edit?usp=sharing) (Google Sheet).