Patent Number: 6,310,199

Title: pH dependent ion exchange matrix and method of use in the isolation of nucleic acids

Abstract: pH dependent ion exchange matrices are provided, with methods for making such matrices, and methods for using such matrices to isolate a target nucleic acid, such as plasmid DNA, chromosomal DNA, or RNA from contaminants, including proteins, lipids, cellular debris, or other nucleic acids. Each pH dependent ion exchange matrix of this invention comprises at least two different ion exchange functional groups, one of which is capable of acting as an anion exchanger at a first pH, and the other of which is capable of acting as a cation exchanger at a second, higher pH. The matrix has an overall neutral charge in a pH range between the first and second pH. The pH dependent ion exchange matrices of the present invention are designed to bind to the target nucleic acid at a pH wherein the overall charge of the matrix is positive, and to release the target nucleic acid as the pH of the surrounding solution is increased. The target nucleic acid can be released from the pH dependent matrix in little or no salt and at about a neutral pH. The matrices and methods of this invention enable one to isolate a target nucleic acid in very few steps, without the use of hazardous chemicals. Target nucleic acids isolated using the pH dependent ion exchange matrices according to the present invention can be used immediately without further extraction or isolation.

Inventors: Smith; Craig E. (Oregon, WI), Holmes; Diana L. (Crystal Lake, IL), Simpson; Daniel J. (Middleton, WI), Katzenhendler; Jehoshua (Jerusalem, IL), Bitner; Rex M. (Cedarburg, WI), Grosch; Josephine C. (Mazomainie, WI)

Assignee: Promega Corporation

International Classification: B01J 41/20 (20060101); B01J 39/26 (20060101); C12N 15/10 (20060101); C07G 003/00 (); C12Q 001/68 (); C07H 021/00 (); C07H 021/02 (); C07H 021/04 ()

Expiration Date: 10/30/2018