Methods and devices for multiplexing amplification reactions
AssigneeApplera Corporation
InventorsJohn C. Gerdes, Elaine Best, Jeffery M. Marmaro
Life of this patent
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Abstract
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 identify E. Coli were identified using the multiplex amplification method.
Claims8
A method for discriminately identifying nucleic acid sequences of bacteria of the coliform species in a sample, comprising:
- (a) contacting said sample with a primer comprising a sequence contained within a sequence identified by base pairs 1733-1900;
- (b) amplifying said nucleic acid sequences;
- and
- (c) detecting said amplified nucleic acid sequences.
A nucleotide sequence containing a primer for discriminately amplifying E. coli bacteria identified by SEQ.ID. NO. 100 or sequence substantially similar to SEQ. ID. NO. 100.
A sequence for discriminately amplifying E. coli bacteria, selected from the group of sequences identified by SEQ. ID. NOS. 85-96.
Dependencies are inferred from the claim text.