Provided herein is a method of detecting short nucleic acids, such as microRNAs, in a sample, such as urine, and a kit for use in detection of the short nucleic acids.

Methods, compositions and kits are described for resequencing, confirming or verifying Next Generation Sequencing (NGS) results with Sanger Sequencing. These methods are particularly useful for samples having very limited quantities such as formalin-fixed, paraffin-embedded (FFPE), Laser Capture Microdissection (LCM), fine needle biopsies or aspirates.

Methods, compositions and kits are described for resequencing, confirming or verifying Next Generation Sequencing (NGS) results with Sanger Sequencing. These methods are particularly useful for samples having very limited quantities such as formalin-fixed, paraffin-embedded (FFPE), Laser Capture Microdissection (LCM), fine needle biopsies or aspirates.

The present invention relates to the amplification and/or detection of nucleic acid molecules. More specifically, the present invention relates to the sensitive amplification, detection, and/or quantification of nucleic acid molecules.

The present invention relates to the amplification and/or detection of nucleic acid molecules. More specifically, the present invention relates to the sensitive amplification, detection, and/or quantification of nucleic acid molecules.

The technology described herein is directed to methods of determining oligonucleotide sequences, e.g. by enriching target sequences prior to sequencing the sequences.

Methods and containers are provided for identifying a species, illustratively a bacterial species. Illustrative methods comprise amplifying various genes in the nucleic acid from the bacterial species in a single reaction mixture using pairs of outer first-stage primers designed to hybridize to generally conserved regions of the respective genes to generate a plurality of first-stage amplicons, dividing the reaction mixture into a plurality of second-stage reactions, each using a unique pair of second-stage primers, each pair of second-stage primers specific for a target bacterial species or subset of bacterial species, detecting which of the second-stage reactions amplified, and identifying the bacterial species based on second-stage amplification. Methods for determining antibiotic resistance are also provided, such methods also using first-stage primers for amplifying genes known to affect antibiotic resistance a plurality of the second-stage reactions wherein each pair of second-stage primers specific for a specific gene for conferring antibiotic resistance.

A method and kit for preparing a high-throughput sequencing library based on nested multiplex PCR are provided. The method includes: amplifying a targeted region using a forward primer and a reverse primer, where the reverse primer includes a reverse specific sequence at a 3′-end and a first universal sequencing sequence at a 5′-end; purifying; and amplifying the purified product using a nested primer located downstream of the forward primer, a first tag primer, and a second universal primer, where the nested primer includes a second universal sequencing sequence at a 5′-end and a specific sequence at a 3′-end, a 3′-end sequence of the first tag primer is partially or completely the same as the first universal sequencing sequence, the first tag primer further includes a first tag sequence, and a 3′-end sequence of the second universal primer is partially or completely the same as the second universal sequencing sequence.

A method and kit for preparing a high-throughput sequencing library based on nested multiplex PCR are provided. The method includes: amplifying a targeted region using a forward primer and a reverse primer, where the reverse primer includes a reverse specific sequence at a 3′-end and a first universal sequencing sequence at a 5′-end; purifying; and amplifying the purified product using a nested primer located downstream of the forward primer, a first tag primer, and a second universal primer, where the nested primer includes a second universal sequencing sequence at a 5′-end and a specific sequence at a 3′-end, a 3′-end sequence of the first tag primer is partially or completely the same as the first universal sequencing sequence, the first tag primer further includes a first tag sequence, and a 3′-end sequence of the second universal primer is partially or completely the same as the second universal sequencing sequence.

According to one embodiment, a detection method is a method for detecting a plurality of target nucleic acids in a sample. The method includes (a) preparing a chain-elongation nucleic acid set group, a primer set, and a probe immobilized substrate, (b) obtaining the target nucleic acid and a long-chain nucleic acid group containing a first sub-chain-elongation nucleic acid and a second sub-chain-elongation nucleic acid, (c) obtaining an amplification product group by maintaining the long-chain nucleic acid group and the primer set under amplification conditions, (d) detecting presence/absence and/or an amount of hybridization, and (e) detecting the plurality of target nucleic acids.