Jan 25, 2016

Public workspaceDNA Extraction Protocol

DOI
dx.doi.org/10.17504/protocols.io.c32yqd
  • Matthew Sullivan1
  • 1Matthew Sullivan Lab, University of Arizona/Ohio State University
  • VERVE Net
  • Sullivan Lab
Verve Team
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DOI: dx.doi.org/10.17504/protocols.io.c32yqd
Protocol Citation: Matthew Sullivan 2016. DNA Extraction Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.c32yqd
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
Created: June 17, 2015
Last Modified: February 17, 2018
Protocol Integer ID: 858
Guidelines
This protocol comes from a group of other protocols. This protocol is (2) of (4):
1. 'Large Volume Marine Cyanophage Phage Protocols'
2. 'DNA Extraction Protocol'
3. 'DNA Precipitation Protocol'
4. 'Checking DNA Concentration with Agarose Gel'

Needed:
  • CsCl
  • Proteinase K
  • SDS
  • Incubator
  • Phenol
  • Microfuge @ 6,000rpm
  • Wide-bore pipette
  • Tube
  • Chloroform
Create mixture of CsCl, Proteinase K, and SDS.
Protocol
CsCl purified phage lysate, Proteinase K, SDS
NAME
CsCl purified phage lysate, Proteinase K, SDS
CREATED BY
Verve Team
Mix 1 volume of dialyzed, CsCl purified phage lysate  with 50 µg/ml Proteinase K
Mix solution in 0.5% SDS.
Mix and incubate 1 hour at 56°C.
Duration01:00:00
Cool to room temperature
Add an equal volume of phenol and invert several times.
Spin 3000g (6000 rpm on microfuge), 5 minutes, room temperature.
Duration00:05:00
Carefully transfer the supernatant with a wide-bore pipette to a fresh tube.
Add an equal volume of phenol:choroform (1:1), invert.
Spin again, 3000g (6000 rpm on microfuge), 5 minutes, room temperature.
Duration00:05:00
Transfer the supernatant, as done before.
Add an equal volume of chloroform and invert.
Spin again, 3000g (6000 rpm on microfuge), 5 minutes, room temperature.
Duration00:05:00
Transfer the supernatant, as done before.
Duration00:05:00