University of Ottawa

Other

Northwestern University

The Perkin-elmer Corporation

Gary W Slater

J William Efcavitch

Roger A O'Neill

Pascal Mayer

Claudia Chiesa

Robert Ruhfel

Hong Yan Zhou

Guy Drouin

Jean Rousseau

Annelise E Barron

Robert Meagher

Laurette C McCormick

Recently two techniques using free solution electrophoresis to separate charged-uncharged polymer conjugates have proven successful: End Labeled Free Solution Electrophoresis (ELFSE) for DNA sequencing, and Free Solution Conjugate Electrophoresis (FSCE) for molar mass profiling of uncharged polymers. Previous attempts have been made to analyze experimental data generated by these new techniques for the electrophoresis of molecules with varying charge distributions. However, the importance of the ends of the polymers in determining the polymer's overall mobility was neglected in previous work. Through a careful investigation and a reanalysis of the experimental data, it is determined here that this 'end effect' critically impacts the behavior of polymers and charged-uncharged polymer conjugates during electrophoresis. In this way, the invention provides for methods that exploit this 'end effect' for the separation of polymeric molecules on the basis of size, including for example DNA separation and sequencing techniques.

Methods for separation of polymeric compounds

An electrophoretic method for purifying a nucleic acid sample is disclosed. According to the method, the electrophoresis is effective to substantially reduce the concentration of contaminants relative to the concentration of desired nucleic acid in the nucleic acid sample, thereby producing a purified nucleic acid. In the method, the loading and recovery wells may be the same or different, and the electric fields may be DC or alternating.

Electrophoretic nucleic acid purification method

An electrophoretic method for purifying a nucleic acid sample is disclosed. The method generally comprises the steps of (1) providing a nucleic acid sample comprising a desired nucleic acid and one or more contaminants, (2) providing an electrophoresis matrix having a loading well and a recovery well formed therein, (3) placing the nucleic acid sample into the loading well, (4) performing a first electrophoresis comprising electrophoresing the nucleic acid sample for a first time effective to transport the desired nucleic acid out of the loading well and into the electrophoresis matrix; and (5) performing a second electrophoresis comprising electrophoresing the nucleic acid sample for a second time effective to transport the desired nucleic acid out of the electrophoresis matrix and into the recovery well. According to the method, the first and second electrophoresis steps are effective to substantially reduce the concentration of contaminants relative to the concentration of desired nucleic acid in the nucleic acid sample, thereby producing a purified nucleic acid. In the method, the loading and recovery wells may be the same or different, and the electric fields may be DC or alternating. Also disclosed is a preparative electrophoresis method employing an alternating electrical field.

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Ottawa, Canada

Gary W Slater