Applera Corporation

Xtrana, Inc.

Applied Biosystems, LLC

Jeffery M Marmaro

John C Gerdes

Jeffrey T Ives

Christopher A Roehl

Elaine A Best

The present invention provides a two-step multiplex amplification reaction wherein the first step truncates the standard initial multiplex amplification round to 'boost' the sample copy number by only a 100-1000 fold increase in the target. Following the first step the product is divided into optimized secondary single amplification reactions, each containing one of the primer sets that were used previously in the first or multiplexed booster step. The booster step can occur using an aqueous target nucleic acid or using a solid phase archived nucleic acid. In particular, nucleic acid sequences that uniquely identifywere identified using the multiplex amplification method.

Methods and devices for multiplexing amplification reactions

This invention is directed to a process for tightly binding nucleic acid to solid phase and corresponding processes for the utilization thereof. Nucleic acid is bound to solid phase matrices exhibiting sufficient hydrophilicity and electropositivity to tightly bind the nucleic acids from a sample. These processes include nucleic acid (double or single stranded DNA and RNA) capture from high volume and/or low concentration specimens, buffer changes, washes, and volume reductions, and enable the interface of solid phase bound nucleic acid with enzyme, hybridization or amplification strategies. The tightly bound nucleic acid may be used, for example, in repeated analyses to confirm results or test additional genes in both research and commercial applications. Further, a method is described for virus extraction, purification, and solid phase amplification from large volume plasma specimens.

Nucleic acid archiving

The present invention provides a two-step multiplex amplification reaction wherein the first step truncates the standard initial multiplex amplification round to 'boost' the sample copy number by only a 100&#x2013;1000 fold increase in the target. Following the first step the product is divided into optimized secondary single amplification reactions, each containing one of the primer sets that were used previously in the first or multiplexed booster step. The booster step can occur using an aqueous target nucleic acid or using a solid phase archived nucleic acid. In particular, nucleic acid sequences that uniquely identifywere identified using the multiplex amplification method.

This invention provides a kit comprising a substrate having a surface coated with a solid phase matrix for nucleic acid manipulation. The solid phase matrix exhibits sufficient hydrophilicity and electropositivity to tightly bind the nucleic acids in a sample. The manipulations include nucleic acid (double or single stranded DNA and RNA) capture from high volume and/or low concentration specimens, buffer changes, washes, and volume reductions, and enable the interface of solid phase bound nucleic acid with enzyme, hybridization or amplification strategies. The tightly bound nucleic acid may be used, for example, in repeated analyses to confirm results or test additional genes in both research and commercial applications. Further, a method for virus extraction, purification, and solid phase amplification from large volume plasma specimens is described.

The present invention pertains to methods, reagents, compositions, kits, and instruments for use in simultaneously amplifying multiple targets. In particular, this invention is based on the discovery of a two-step multiplex amplification reaction wherein the first step truncates the standard initial multiplex amplification round to &ldquo;boost&rdquo; the sample copy number by only a 100-1000 fold increase in the target. Following the first step the product is divided into optimized secondary single amplification reactions, each containing one of the primer sets that were used previously in the first or multiplexed booster step. The booster step can occur using an aqueous target nucleic acid or using a solid phase archived nucleic acid.

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Aurora, CO, United States of America

Jeffery M Marmaro