The Perkin-elmer Corporation

Pe Corporation (ny)

Other

Applera Corporation

University of Ottawa

Claudia Chiesa

Roger A O'Neill

Jer-Kang Chen

J William Efcavitch

Pascal Mayer

Stephen E Moring

David M Demorest

George A Fry

Robert Ruhfel

Gary W Slater

Hong Yan Zhou

Guy Drouin

Jean Rousseau

Michael Egholm

Majid Mehrpouyan

Stephen Lambert

Vergine C Furniss

Gary Schroth

George Fry

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

The present invention provides methods and kits for isolating one strand of a double-stranded target nucleic acid. The method capitalizes on the differences in the kinetics and thermodynamic stabilities between conventional DNA/DNA, DNA/RNA and RNA/RNA duplexes and heteroduplexes in which one strand of the heteroduplexe is a nucleobase polymer having a net positively charged or net neutral backbone, such as a PNA.

Strand displacement methods employing competitor oligonucleotides for isolating one strand of a double-stranded nucleic acid

The present invention provides substrates and apparatuses for efficient, rapid and specific capture, and optimal recovery, of nucleic acids, as well as methods of their use. The substrate is porous in nature and has a capture polynucleotide capable of hybridizing to a target nucleic acid immobilized thereon. Upon flowing a sample containing or suspected of containing the target nucleic acid through the porous substrate, the target nucleic acid is rapidly captured. Following capture, the target nucleic acid can be efficiently recovered for subsequent use.

Method and apparatus for flow-through hybridization

The invention relates to methods and compositions for simultaneously generating a plurality of polynucleotide sequencing ladders or PCR amplification products. Each sequencing ladder is generated from a recoverable primer, i.e., an oligonucleotide primer comprising a recovery tag. The recovery tag may be an oligonucleotide. Each sequencing ladder has a unique recovery tag. After the generation of the multiple sequencing ladders, the different sequencing ladders are separated from one another, i.e., purified, by binding to recovery tag binding compounds that have been immobilized on one or more solid supports. The recovery tag binding compounds are immobilized on the solid support in an addressable manner, i.e., the recovery tag binding compounds have distinct locations on the solid support. The binding of the sequencing ladders to the recovery tag binding compounds serves to separate the different polynucleotide sequencing ladders present in a given solution. The separated sequencing ladders may then be released from the immobilized recovery tag binding compounds and subsequently resolved into individual components of the sequencing ladders or PCR products. The subject methods of separating sequencing ladders simultaneously generated in the same vessel may readily be adapted to separate a plurality of simultaneously generated polynucleotide amplification products. Other aspects of the invention are kits for the generation or recovery of a plurality of polynucleotide sequencing ladders or amplification products. The kits comprise a plurality of recoverable primers having unique recovery tags. Preferably, the recovery tags are polynucleotides that have substantially the same denaturation temperature when bound to appropriate recovery tag binding compounds, i.e., the primers comprise an integrated set. The kits may further comprise recovery tag binding compounds. The kits may further comprise labeled chain terminators. Other aspects of the invention include polynucleotide recovery devices.

Multiplex polynucleotide capture methods and compositions

Disclosed is a method for increasing the electroosmotic flow rate available for a silica surface. In the method, there is provided an electrophoretic channel which is defined by one or more silica surfaces. The surface(s) are contacted with an alkaline aqueous solution containing a solubilized silicate-monovalent metal complex in an amount effective to increase the acidity of the silica surface(s), as evidenced by a reduction in the average bulk pKa of the surface(s). The achieved increase in acidity is greater than would be obtained using an otherwise identical solution lacking said silicate. In one preferred embodiment, the monovalent metal used in the solution is Li,, Na,, or K,. Also disclosed is a method for increasing the acidity of a silica surface, by contacting the surface with an alkaline aqueous solution of the type noted above.

Method and silicate composition for conditioning silica surfaces

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.

Disclosed is a method for increasing the electro-osmotic flow rate available for a silica surface. In the method, there is provided an electrophoretic channel which is defined by one or more silica surfaces. The surface(s) are contacted with an alkaline aqueous solution containing a solubilized silicate-monovalent metal complex in an amount effective to increase the acidity of the silica surface(s), as evidenced by a reduction in the average bulk pKa of the surface(s). The achieved increase in acidity is greater than would be obtained using an otherwise identical solution lacking said silicate. In one preferred embodiment, the monovalent metal used in the solution is Li.sup.+, Na.sup.+, or K.sup.+. Also disclosed is a method for increasing the acidity of a silica surface, by contacting the surface with an alkaline aqueous solution of the type noted above.

Growing community of inventors

Foster City, CA, United States of America

Claudia Chiesa