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Method of plasmid DNA production and purification

  • US 5,981,735 A
  • Filed: 02/12/1997
  • Issued: 11/09/1999
  • Est. Priority Date: 02/12/1996
  • Status: Expired due to Fees
First Claim
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1. A method for isolation and purification of plasmid DNA, comprising the steps of:

  • a) determining the irreversible alkaline denaturation value of said plasmid DNA;

    b) growing a culture of bacterial host cells to a cell density within the range of about 2 g to about 200 g per liter dry weight units, wherein said host cells contain plasmid DNA having an irreversible alkaline denaturation value;

    c) lysing said bacterial cells by contacting said culture during exponential growth with an amount of alkali sufficient to reach a pH value that is about 0.1 pH unit to about 0.2 pH unit below the irreversible alkaline denaturation value of said plasmid DNA; and

    d) isolating a said plasmid DNA preparation.

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