Purification and use of gellan in electrophoresis gels
AssigneeShin-Etsu Chemical Co., Ltd.
InventorsLinda Thorne, Richard W. Armentrout
Life of this patent
Estimating the term…
Abstract
Gellan can be purified from nucleic acid contamination by combining the contaminated gellan with DNase under conditions that allow the DNase to degrade the nucleic acid contaminant. The purified gellan is useful in gel electrophoresis. A buffer which allows cystamine to be used as a reversible cross-linker does not have to be recirculated during the course of a normal gel run.
Claims54
A method for purifying gellan, comprising:
- (a) combining DNase and gellan, the gellan being contaminated with nucleic acid, thereby providing a mixture;
- and
- (b) maintaining the mixture of step (a) under conditions where the DNase degrades at least some of the nucleic acid, thereby providing purified gellan.
A gellan composition comprising water and gellan, the composition containing either no nucleic acid or nucleic acid at a concentration of less than 10 ppm based on the weight of the gellan.
A composition suitable for use in preparing an electrophoresis medium, comprising:
- (a) gellan;
- and
- (b) either no nucleic acid or nucleic acid at a concentration of less than 10 ppm nucleic acid, based on the weight of gellan.
A kit comprising:
- (a) a matrix composition comprising gellan and nucleic acid at a concentration of less than 10 ppm based on the weight of the gellan;
- (b) buffer;
- and
- (c) cross linking agent.
A method of performing electrophoresis comprising
- (1) forming an electrophoresis medium by combining ingredients comprising:
- (a) a matrix composition comprising gellan, nucleic acid at a concentration of less than 10 ppm based on the weight of the gellan, and size-separation property modifying polymer;
- (b) buffer;
- and
- (c) cross linking agent;
- and
- (2) applying an electric field across the medium.
An electrophoresis apparatus comprising:
- (a) a cross linked matrix formed by combining gellan, cross linking agent, nucleic acid at a concentration of less than 10 ppm based on the weight of the gellan, buffer, and size-separation property modifying polymer;
- and
- (b) an apparatus for exposing said cross linked matrix to an electric field.
A method for recovering a biological material, comprising:
- (a) adding a mixture comprising a biological material to a cross linked electrophoresis medium, the medium being formed by a method comprising combining a cross linking agent and gellan contaminated with less than 10 ppm nucleic acid based on the weight of the gellan;
- (b) exposing the medium to an electric field to separate in said medium said biological material from other components in the mixture;
- (c) removing a zone of the medium containing the biological material from the medium;
- (d) exposing the removed zone to an agent that reverses the cross linking of the medium, to provide liquefied electrophoresis medium;
- and
- (e) separating the biological material from the liquefied electrophoresis medium, thereby recovering the biological material.
A composition comprising water, imidazole or a salt thereof, and boric acid or a salt thereof.
Dependencies are inferred from the claim text.