Antonio O Reyes

US Patent:

6335184, Jan 1, 2002

Filed:

Jan 11, 1999

Appl. No.:

09/228324

Inventors:

Antonio Arevalo Reyes - Danville CA
Robert Bruce Wallace - Greenbrae CA
Luis A. Ugozzoli - San Rafael CA

Assignee:

Bio-Rad Laboratories, Inc. - Hercules CA

International Classification:

C12P 1934

US Classification:

435 912, 435 6

Abstract:

The extensive synthesis (âamplificationâ) of a nucleic acid sequence of interest is attained through a linked series of multi-cycle primer extension reactions (LLA). The primers used in each of the primer extension reactions of the process contain non-replicable and/or cleavable elements that halt nucleic acid synthesis and thereby prevent the synthesized molecules from serving as templates in subsequent cycles. Synthesized molecules accumulate during primer extension in a mathematically linear fashion, thereby rendering the process relatively insensitive to contaminating nucleic acids. Multiple primer sets are employed, and simultaneously included in the reaction mixture, thereby ensuring the accumulation of a large number of copies of the nucleic acid sequence of interest. The invention also provides for the detection of an amplified nucleic acid sequence of interest, as well as reagent kits for carrying out the reaction.

US Patent:

20040091923, May 13, 2004

Filed:

Oct 6, 2003

Appl. No.:

10/680341

Inventors:

Antonio Reyes - Danville CA, US
Robert Wallace - Greenbrae CA, US
Luis Ugozzoli - San Rafael CA, US

Assignee:

Bio-Rad Laboratories, Inc. - Hercules CA

International Classification:

C12Q001/68
C12P019/34

US Classification:

435/006000, 435/091200

Abstract:

The generation and extensive synthesis (“amplification”) of a cDNA sequence of interest is attained through a linked series of multi-cycle primer extension reactions (RT-LLA). The primers used in each of the primer extension reactions of the process contain non-replicable and, in some cases non-replicable and cleavable, elements that halt nucleic acid synthesis and thereby prevent the synthesized molecules from serving as templates in subsequent cycles. Synthesized molecules accumulate during primer extension in a mathematically linear fashion, thereby rendering the process relatively insensitive to contaminating nucleic acids. Multiple primer sets are employed, and simultaneously included in the reaction mixture, thereby ensuring the accumulation of a large number of copies of the cDNA sequence of interest.