Brandeis University

J Aquiles Sanchez

Lawrence J Wangh

John E Rice

Kenneth E Pierce

Cristina Hartshorn

Arthur Henry Reis, Jr

Jesse J Salk

Arthur Reis

Yanwei Jia

Skye Fishbein

Stephen Van Hooser

Jr Arthur H Reis

Kenneth Pierce

Provided herein are reagents for improving PCR accuracy.

Reagents for improving PCR accuracy

Provided herein are kits for performing for nucleic acid sequences.

Kits and reaction mixtures for analyzing single-stranded nucleic acid sequences

Homogenous detection during or following PCR amplification, preferably LATE-PCR, utilizing fluorescent DNA dye and indirectly excitable labeled primers and probes, improves reproducibility and quantification. Low-temperature homogeneous detection during or following non-symmetric PCR amplification, preferably LATE-PCR, utilizing fluorescent DNA dye and indirectly excitable labeled mismatch-tolerant probes permits analysis of complex targets. Sequencing sample preparation methods following LATE-PCR amplifications reduce complexity and permit 'single-tube' processing.

Methods for sequential DNA amplification and sequencing

A non-symmetric polymerize chain reaction (PCR) amplification method employing a limiting primer in low concentration whose concentration-adjusted melting point at least equals, and preferably exceeds, that of the excess primer, the latter in turn not being more than 25&#xb0; C. below the melting temperature of the amplicon. Assays employing such amplification and labeled hybridization probes, including assays that include a detection step following primer extension or a low-temperature probe, or both. Kits for performing such assays and primer or primer-and-probe sets for performing the foregoing amplifications and assays.

Late-PCR

Provided herein are fluorescence detection methods for nucleic acid sequences and to kits for performing such methods.

Methods, kits and reaction mixtures for analyzing single-stranded nucleic acid sequences

This disclosure relates to amplification and detection of a rare variant or variants of a DNA sequence in an abundant variant of the sequence, such as detection of a low-level somatic mutations and minority alleles in an excess of normal nucleic acid target sequences.

Detecting nucleic acid variations within populations of genomes

A non-symmetric polymerise chain reaction (PCR) amplification method employing a limiting primer in low concentration whose concentration-adjusted melting point at least equals, and preferably exceeds, that of the excess primer, the latter in turn not being more than 25&#xb0; C. below the melting temperature of the amplicon. Assays employing such amplification and labeled hybridization probes, including assays that include a detection step following primer extension or a low-temperature probe, or both. Kits for performing such assays and primer or primer-and-probe sets for performing the foregoing amplifications and assays.

An assay comprising more than one primer pair and more than one detection probe, a low copy number synthetic amplicon corresponding to each of the primer pairs. The assay can detect and distinguish between various sub-types and strains of an influenza virus using any suitable nucleic acid amplification technique. Related kits and methods are also described.

Detection and analysis of influenza virus

Primers, probes and methods for nucleic acid amplification

An additive for preventing mispriming in polymerase chain reaction (PCR) amplifications and assays comprising a hairpin oligonucleotide having a stem duplex greater than six nucleotides in length and a stabilized stem terminus. The additive improves PCR amplifications, including LATE-PCR amplifications when added to initial amplification reaction mixtures. It can be included in oligonucleotide sets and in kits for PCR amplification and assays.

Reagents and methods for improving reproducibility and reducing mispriming in PCR amplification

A non-symmetric polymerase chain reaction (PCR) amplification method employing a limiting primer in low concentration whose concentration-adjusted melting point at least equals, and preferably exceeds, that of the excess primer, the latter in turn not being more than 25&#xb0; C. below the melting temperature of the amplicon. Assays employing such amplification and labeled hybridization probes, including assays that include a detection step following primer extension or a low-temperature probe, or both. Kits for performing such assays and primer or primer-and-probe sets for performing the foregoing amplifications and assays.

Growing community of inventors

Framingham, MA, United States of America

J Aquiles Sanchez