Deborah D Dodge

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.