The present invention relates to a method for identifying a microorganism in a biological sample by polymerase chain reaction (PCR), comprising the steps of a) providing a biological sample suspected of comprising microbes, and optionally isolating nucleic acid sequences from said biological sample; b) PCR amplifying at least one microbial rRNA internal transcribed spacer (ITS) region comprised in said optionally isolated nucleic acid sequences using a set of broad-taxonomic range amplification primers to thereby generate PCR amplicons from nucleic acid sequences of microbial origin; c) recording a high resolution melting curve for the PCR amplicons, and recording the length of the PCR amplicons; d) comparing the high resolution melting curve with a database comprising high resolution melting curves of reference amplicons of known microbial species or strains, to thereby obtain a first identity indicator; e) comparing the length of each PCR amplicon having a distinct length with a database comprising PCR amplicon lengths of reference amplicons of known microbial species or strains, to thereby obtain a second identity indicator; and f) identifying the microorganism present in said sample to the species or strain level if the first and second identity indicator match.

Described herein are compositions, methods, and techniques to generate a cancer signature and uses thereof. The cancer signature can be used to determine a cancer progression risk of a subject based upon expression levels of genes of a progression gene signature in a sample. The methods can be used to predict a prognosis, to select an appropriate treatment regimen, to identify or screen for an agent effective against a cancer, or a combination thereof. Computer implemented methods and systems that implement those methods are also provided. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present disclosure.

Described herein are compositions, methods, and techniques to generate a cancer signature and uses thereof. The cancer signature can be used to determine a cancer progression risk of a subject based upon expression levels of genes of a progression gene signature in a sample. The methods can be used to predict a prognosis, to select an appropriate treatment regimen, to identify or screen for an agent effective against a cancer, or a combination thereof. Computer implemented methods and systems that implement those methods are also provided. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present disclosure.

Compositions, kits, and methods for performing rapid polymerase chain reaction (PCR) to amplify a target nucleic acid in a biological sample are disclosed. The methods include the use of at least one hybridization stabilizer and/or the adjustment of the thermocycling profiles between initiation and propagation phases of the amplification process. Also disclosed are methods of detecting the target nucleic acid following amplification thereof, as well as reaction mixtures that may be utilized in said methods.

Compositions, kits, and methods for performing rapid polymerase chain reaction (PCR) to amplify a target nucleic acid in a biological sample are disclosed. The methods include the use of at least one hybridization stabilizer and/or the adjustment of the thermocycling profiles between initiation and propagation phases of the amplification process. Also disclosed are methods of detecting the target nucleic acid following amplification thereof, as well as reaction mixtures that may be utilized in said methods.

Compositions, kits, and methods for performing rapid polymerase chain reaction (PCR) to amplify a target nucleic acid in a biological sample are disclosed. The methods include the use of at least one hybridization stabilizer and/or the adjustment of the thermocycling profiles between initiation and propagation phases of the amplification process. Also disclosed are methods of detecting the target nucleic acid following amplification thereof, as well as reaction mixtures that may be utilized in said methods.

The present disclosure generally relates, in certain aspects, to droplet-based microfluidic devices and methods. In certain aspects, target nucleic acids contained within droplets are amplified within droplets in a first step, where multiple primers may be present. However, multiple primers may cause multiple target nucleic acids to be amplified within the droplets, which can make it difficult to identify which nucleic acids were amplified. In a second step, the amplified nucleic acids may be determined. For example, the droplets may be broken and the amplified nucleic acids can be pooled together and sequenced. As an example, new droplets may be formed containing the amplified nucleic acids, and those nucleic acids within the droplets amplified by exposure to certain primers.

The present disclosure generally relates, in certain aspects, to droplet-based microfluidic devices and methods. In certain aspects, target nucleic acids contained within droplets are amplified within droplets in a first step, where multiple primers may be present. However, multiple primers may cause multiple target nucleic acids to be amplified within the droplets, which can make it difficult to identify which nucleic acids were amplified. In a second step, the amplified nucleic acids may be determined. For example, the droplets may be broken and the amplified nucleic acids can be pooled together and sequenced. As an example, new droplets may be formed containing the amplified nucleic acids, and those nucleic acids within the droplets amplified by exposure to certain primers.

The present disclosure generally relates, in certain aspects, to droplet-based microfluidic devices and methods. In certain aspects, target nucleic acids contained within droplets are amplified within droplets in a first step, where multiple primers may be present. However, multiple primers may cause multiple target nucleic acids to be amplified within the droplets, which can make it difficult to identify which nucleic acids were amplified. In a second step, the amplified nucleic acids may be determined. For example, the droplets may be broken and the amplified nucleic acids can be pooled together and sequenced. As an example, new droplets may be formed containing the amplified nucleic acids, and those nucleic acids within the droplets amplified by exposure to certain primers.

The present invention provides a simple and highly accurate method for an ecological survey with low environmental burden, more specifically, a method for analyzing biological species living in a water environment. The method assesses biological species living in the water environment quantitatively and further identify them exhaustively by analyzing RNA contained in the water environment.