Nicole Hampton Ellis

US Patent:

60430702, Mar 28, 2000

Filed:

Aug 29, 1997

Appl. No.:

8/924031

Inventors:

Nicole M. Ellis - San Mateo CA
Robert G. Kuimelis - Redwood City CA
Cheryl R. Heiner - La Honda CA
Katherine D. Lazaruk - Redwood City CA
Patric Sean Walsh - Danville CA

Assignee:

The Perkin-Elmer Corporation

International Classification:

C12N 910
C07H 2104

US Classification:

435193

Abstract:

The invention relates to novel chimeric phosphoramidate oligonucleotides and their use in primer-extension methods such as DNA sequencing and nucleic acid amplification. The subject chimeric phosphoramidate oligonucleotides have both N3'-phosphoramidate linkages and phosphodiester linkages. The invention includes methods of primer extension using the subject chimeric oligonucleotides as primers. Primer extension methods of interest include nucleic acid amplification reactions, e. g. PCR, and polynucleotide sequencing reactions. In the primer extension methods of the invention, a chimeric phosphoramidate oligonucleotide primer is annealed to a polynucleotide template. After annealing, the chimeric oligonucleotide primer is extended by joining a nucleotide to the 3' end of the primer by a DNA polymerase catalyzed reaction. Other embodiments of the invention include methods of primer extension using phosphoramidate linkage containing polynucleotide templates. Such methods include annealing an oligonucleotide primer (chimeric or otherwise) to a polynucleotide template comprising at least one phosphoramidate linkage.

US Patent:

60308133, Feb 29, 2000

Filed:

Mar 16, 1999

Appl. No.:

9/271060

Inventors:

Nicole M. Ellis - San Mateo CA
Robert G. Kuimelis - Redwood City CA
Cheryl R. Heiner - La Honda CA
Katherine D. Lazaruk - Redwood City CA
Patric Sean Walsh - Danville CA

Assignee:

The Perkin-Elmar Corporation - Foster City CA

International Classification:

C12P 1934

US Classification:

435 911

Abstract:

The invention relates to novel chimeric phosphoramidate oligonucleotides and their use in primer-extension methods such as DNA sequencing and nucleic acid amplification. The subject chimeric phosphoramidate oligonucleotides have both N3'-phosphoramidate linkages and phosphodiester linkages. The invention includes methods of primer extension using the subject chimeric oligonucleotides as primers. Primer extension methods of interest include nucleic acid amplification reactions, e. g. PCR, and polynucleotide sequencing reactions. In the primer extension methods of the invention, a chimeric phosphoramidate oligonucleotide primer is annealed to a polynucleotide template. After annealing, the chimeric oligonucleotide primer is extended by joining a nucleotide to the 3' end of the primer by a DNA polymerase catalyzed reaction. Other embodiments of the invention include methods of primer extension using phosphoramidate linkage containing polynucleotide templates. Such methods include annealing an oligonucleotide primer (chimeric or otherwise) to a polynucleotide template comprising at least one phosphoramidate linkage.

US Patent:

59981431, Dec 7, 1999

Filed:

Dec 5, 1997

Appl. No.:

8/986176

Inventors:

Nicole M. Ellis - San Mateo CA
Deborah E. Dodge - Albany CA
Douglas H. Smith - Centerville DE

Assignee:

The Perkin-Elmer Corporation - Foster City CA

International Classification:

C12Q 168
C12P 1934
C07H 2102
C07H 2104

US Classification:

435 6

Abstract:

The present invention relates to improved methods of generating polynucleotide sequencing reaction products from cycle sequencing and relates to various instruments and reagents for use in the subject methods. The use of "step-down" thermal profiles in conjunction with cycle sequencing is described. Step-down thermal profiles are formed by combining several thermal cycle sets such that the annealing temperature of each thermal cycle set is less than the annealing temperature of the preceding thermal cycle set. One embodiment of the invention is a method of generating a plurality of polynucleotide sequencing reaction products in parallel by subjecting a plurality of sequencing solution preparations to a step-down thermal profile, i. e. , exposure to repeated thermal cycle sets, each thermal cycle set having an annealing temperature that is lower than the annealing temperature of the annealing phases of the preceding thermal cycle set. Other embodiments of the invention include systems for generating a plurality of polynucleotide sequencing reaction products in parallel.