Diverse cDNA libraries derived from cell-free mRNA and methods of preparing the same are provided. The library may be prepared by extracting RNA from a bodily fluid such as serum or plasma, separating the RNA from contaminants, synthesizing cDNA with a reverse transcriptase enzyme, and enriching protein-coding nucleotide sequences. The library may include a multiplicity of transcripts from solid tissues. Cf-RNA can be measured by qPCR, sequencing, or other suitable methods.

The application provides an agnostic, shotgun nucleic acid sequencing-based method for the detection of pathogens in samples from human patients, animals, or plants. The method includes dehosting the sample of the nucleic acid molecules of host origin and provides for the detection of pathogens without prior knowledge of their genome sequences.

An approach for differentially isolating eukaryotic (plant and animal) DNA from bacterial DNA prior to sequencing using a combination of size exclusion-based separation and differential cell lysis. The method of the present invention exploits the differences of the cellular size and components of each type of organism to be separated. The composition and nature of the cell wall of plant cells, enzymatic sensitivity of bacterial and animal cells and overall size difference of bacterial and plant/animal cells allows one portion of a mixed sample to be lysed while retaining the integrity of the remaining organisms. Separation of one phylogenetic component then permits the remaining components to be extracted with minimal contribution from the preceding component. The separation of DNAs from differing contributing kingdoms in an unknown sample increases interpretability through decreasing complexity in subsequent sequencing applications.

An approach for differentially isolating eukaryotic (plant and animal) DNA from bacterial DNA prior to sequencing using a combination of size exclusion-based separation and differential cell lysis. The method of the present invention exploits the differences of the cellular size and components of each type of organism to be separated. The composition and nature of the cell wall of plant cells, enzymatic sensitivity of bacterial and animal cells and overall size difference of bacterial and plant/animal cells allows one portion of a mixed sample to be lysed while retaining the integrity of the remaining organisms. Separation of one phylogenetic component then permits the remaining components to be extracted with minimal contribution from the preceding component. The separation of DNAs from differing contributing kingdoms in an unknown sample increases interpretability through decreasing complexity in subsequent sequencing applications.

Systems and methods for performing a plurality of nucleic acid amplification assays in an automated analyzer. A first nucleic acid amplification assay of the plurality is performed in accordance with a first set of assay parameters which consist of system-defined parameters. And a second nucleic acid amplification assay of the plurality is performed in accordance with a second set of assay parameters which includes one or more user-defined parameters.

Disclosed are a method for DNA extraction in a sample for next generation sequencing (NGS) and a method of constructing a NGS library using the extracted DNA. The method for DNA extraction includes: preparing a mixture by mixing a biological sample with a buffer; applying microwaves to the mixture; and recovering DNA. The method of constructing a NGS library includes: extracting DNA according to the method for DNA extraction; amplifying a target DNA using primers; and purifying the amplified product and subjecting the purified product to library pooling.

Disclosed are a method for DNA extraction in a sample for next generation sequencing (NGS) and a method of constructing a NGS library using the extracted DNA. The method for DNA extraction includes: preparing a mixture by mixing a biological sample with a buffer; applying microwaves to the mixture; and recovering DNA. The method of constructing a NGS library includes: extracting DNA according to the method for DNA extraction; amplifying a target DNA using primers; and purifying the amplified product and subjecting the purified product to library pooling.

The present invention relates to methods, kits and uses for diagnosing heart failure using miRNA-biomarker from blood.

The present invention relates to methods, kits and uses for diagnosing heart failure using miRNA-biomarker from blood.

Methods of isolating nucleic acids comprising DNA from biological material are disclosed. The methods comprise a lysis step using an aqueous composition which comprises lithium at a concentration of 0.05 to 1.0M, a chelating agent, and a surfactant to produce a lysed composition. The lysed composition is treated with a solid support that is capable of immobilising DNA in the presence of a dissolved chaotropic agent at a concentration of 0.05 to 2M and 25% to 60% by volume of a C.sub.1-3 alkanol. The support is then washed with a first wash solution containing lithium dissolved in a C.sub.1-3 alkanol. Subsequently, the support is washed with a liquid comprising at least 80% by volume of a C.sub.1-3 alkanol. The nucleic acid comprising DNA is then eluted from the support.