BioTechniques (Mar 2002)

High-Speed Plasmid Isolation Using 96-Well, Size-Exclusion Filter Plates

  • D. Harris,
  • M. Engelstein,
  • R. Parry,
  • J. Smith,
  • M. Mabuchi,
  • J. Leonard

DOI
https://doi.org/10.2144/02323pf02
Journal volume & issue
Vol. 32, no. 3
pp. 626 – 631

Abstract

Read online

The accelerating pace of genomics analysis has necessitated the abbreviation of DNA sample preparation protocols. We have developed a size-exclusion-based system for the rapid isolation of plasmid DNA in a 96-well microplate format. This high-speed protocol employs a modified alkaline lysis method for the preparation of the bacterial lysate, followed by three short vacuum filtration steps. Unlike traditional bind/wash/elute methods, there is no need to use chaotropic salts or ethanol. The samples are recovered from the top side of the MultiScreen96® PLASMID plates. Starting with bacterial cell pellets, the entire procedure for purifying the plasmid DNA can be performed in 30 min with a multichannel pipettor. The high yields, reproducibility, and quality of the plasmids make this system a good choice for any cloning or DNA sequencing operation.