High-speed plasmid isolation using 96-well, size-exclusion filter plates

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

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)626-631
Number of pages6
JournalBioTechniques
Volume32
Issue number3
StatePublished - Mar 20 2002
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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    Harris, D., Engelstein, M., Parry, R., Smith, J., Mabuchi, M., & Leonard, J. (2002). High-speed plasmid isolation using 96-well, size-exclusion filter plates. BioTechniques, 32(3), 626-631.