---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2009-01-06 23:09:06+00:00
layout: post
slug: pcr-dungan-isolates-2
title: PCR - Dungan Isolates
categories:
  - 2009
  - Miscellaneous
tags:
  - BC05Ca-18t/H5
  - Chelex
  - DNA Isolation
  - Dungan isolates
  - gel
  - PCR
  - VATm-1.2t
  - VNTc-1.5t
  - VNTc-12-C1/G10
  - xCvC-11t
  - xCvC-12t
  - xCvC-17t
---

All samples , except xCVC-11t, are already in Chelex. For xCvC-11t, pipetted a shunk of cells/tissue from source tube. Volume of liquid (EtOH) was ~75uL. Added this to screw cap tube containing 300uL of 10% Chelex solution. Vortexed and incubated @ 95C for 30mins. Vortexed and incubated other samples at 95C for 5mins. Set up PCR with AmpliTaq. Anneal temp. = 53C. [PCR set up here](https://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20090107-01.jpg).

![](https://eagle.fish.washington.edu/Arabidopsis/20090106.png)

Lane 1 - 100bp ladder

Lane 2 - xCvC-11t

Lane 3 - xCvC-12t

Lane 4 - xCvC-17t

Lane 5 - VNTc-12-C1/G10

Lane 6 - BC05Ca-18t/H5

Lane 7 - VATm-1.2t

Lane 8 - VNTc-1.5t

Lane 9 - Neg. Control

Results: Ladder is degraded and there are no bands in any lane. Will repeat and try to duplicate Steven's results from 20080916.
