title: High-throughput plasmid mini preparations facilitated by micro-mixing.
abstract: We have developed a reliable high-throughput plasmid isolation system using a 96-well plate format. This system combines a novel glass bead micro-mixing method with modified alkaline lysis and Sephacryl S-500 DNA purification procedures. Mechanical forces generated by vortexing glass beads inside each well of the 96-well plates ensure that the bacterial pellets are homogeneously resuspended, the cells are completely lyzed, and the resulting bacterial lysates are thoroughly mixed with the potassium acetate solution. The vortexing speed and duration for glass bead mixing have been standardized to facilitate plasmid DNA yields without significant adjustments.
authors.affiliation: Department of Molecular Biotechnology, University of Washington, Seattle 98195, USA. vng@u.washington.edu
authors.forename: W L
authors.initials: WL
authors.lastname: Ng
authors.forename: M
authors.initials: M
authors.lastname: Schummer
authors.forename: F D
authors.initials: FD
authors.lastname: Cirisano
authors.forename: R L
authors.initials: RL
authors.lastname: Baldwin
authors.forename: B Y
authors.initials: BY
authors.lastname: Karlan
authors.forename: L
authors.initials: L
authors.lastname: Hood
mesh_terms: D002478:Cells, Cultured; D004274:DNA, Recombinant / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D010051:Ovarian Neoplasms / Q000235:genetics / Q000473:pathology; D010053:Ovary / Q000166:cytology / Q000378:metabolism; D010957:Plasmids; D014407:Tumor Cells, Cultured
chemical_list: D004274:DNA, Recombinant