Purification of plasmid DNA using a multicompartment electrolyser separated by ultrafilter membranes View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2002-01

AUTHORS

Zhijun Wang, Guowei Le, Yonghui Shi, Grzegorz Węgrzyn

ABSTRACT

A simple, scalable method for purification of plasmid DNA is described. Plasmid DNA was released from Escherichia coli JM109 by lysis (1% SDS, 0.2 M NaOH). Then a neutralization solution (3 M sodium acetate buffer, pH 4.8) was added to precipitate genomic DNA and protein. After the clarification of the lysate, the supernatant was placed in a multicompartment electrolyser separated by ultrafilter membranes to remove the remaining contamination (RNA, genomic DNA and protein). A recovery of 75%±2% of total plasmid DNA was obtained after 60 min electrophoresis with a field strength of 8 V cm−1 using cells at 30 g l−1 (quantified by dry cell weight). Genomic DNA, RNA and protein were undetectable in the purified plasmid DNA solution. More... »

PAGES

121-124

References to SciGraph publications

  • 1997-02. The processing of a plasmid-based gene from E. Coli. Primary recovery by filtration in BIOPROCESS AND BIOSYSTEMS ENGINEERING
  • 1999-01. Effect of shear on plasmid DNA in solution in BIOPROCESS AND BIOSYSTEMS ENGINEERING
  • Journal

    TITLE

    Biotechnology Letters

    ISSUE

    2

    VOLUME

    24

    Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1023/a:1013839819916

    DOI

    http://dx.doi.org/10.1023/a:1013839819916

    DIMENSIONS

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