Nov 05, 2024

Public workspacePlasmid Miniprep with ethanol precipitation

 Forked from Plasmid Miniprep
This protocol is a draft, published without a DOI.
  • 1University of Washington
Icon indicating open access to content
QR code linking to this content
Protocol CitationMatthew Akamatsu 2024. Plasmid Miniprep with ethanol precipitation. Protocol exchange https://protocols.io/view/plasmid-miniprep-with-ethanol-precipitation-dq7e5zje
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
We use this protocol and it's working
Created: November 05, 2024
Last Modified: November 05, 2024
Protocol Integer ID: 111558
Abstract
Northeastern's method for plasmid extraction and purification. From Thermo Scientific's GeneJET Plasmid Miniprep Kit

Pellet culture in a 1.8mL centrifuge tube at 8,000rpm for 1 minute. Decant Media
Add 250uL of Resuspension Solution and vortex
Add 250uL of lysis solution and invert 4-6 times
Add 350uL of Neutralization Solution and invert the tube 4-6 times
Add 1x volume of 96% ethanol
Bind the DNA to the column by decanting
Centrifuge at 13,300 rpm for 1 minute
Duration00:01:00
Wash the column with 500uL of wash solution and centrifuge for 1 minute
Duration00:01:00
Discard the flow-thru and centrifuge for 1 minute
Duration00:01:00
Elute the purified DNA with 50uL of Elution Buffer. Let incubate for two minutes, then centrifuge for 2 minutes. Collect the flow-thru in a new centrifuge tube
Duration00:01:00