Provided herein is a method for detecting the presence of a COVID-19 virus RNA or other pathogenic respiratory viruses, such as an influenza virus, or other RNA of interest in a sample. Nucleic acids are obtained from the sample and are used as a template in a combined isothermal reverse transcription, RNAse H and isothermal amplification reaction to generate single stranded RNA amplicons containing sequences complementary to fluorescent labeled detector probes. The single-stranded RNA amplicons hybridize to the detector probe and to hybridization probes with sequences complementary to a sequence determinant in the COVID-19 or other virus RNAs. The microarray is imaged to detect fluorescent signals thereby identifying the virus.

Provided is a method for identifying whether an RNA molecule has a 2′-O-methylation modification on a nucleotide, said method comprising: (1) contacting the RNA molecule with a ribonuclease Rnase R; and (2) detecting whether the RNA molecule is degraded or detecting hydrolysis termination positions after degradation. If the RNA molecule is degraded, this indicates that the RNA molecule does not have a 2′-O-methylation modification on a 3′ terminal nucleotide, and if hydrolysis terminates at a same site on multiple random broken fragments, this indicates that the RNA molecule has a 2′-O-methylation modification on the nucleotide at the position immediately preceding the termination site. Also provided are applications of the method for screening for a disease diagnosis target and confirming whether a subject has a 2′-O-methylation modification-related disease.

Provided is a method for identifying whether an RNA molecule has a 2′-O-methylation modification on a nucleotide, said method comprising: (1) contacting the RNA molecule with a ribonuclease Rnase R; and (2) detecting whether the RNA molecule is degraded or detecting hydrolysis termination positions after degradation. If the RNA molecule is degraded, this indicates that the RNA molecule does not have a 2′-O-methylation modification on a 3′ terminal nucleotide, and if hydrolysis terminates at a same site on multiple random broken fragments, this indicates that the RNA molecule has a 2′-O-methylation modification on the nucleotide at the position immediately preceding the termination site. Also provided are applications of the method for screening for a disease diagnosis target and confirming whether a subject has a 2′-O-methylation modification-related disease.

The present invention relates to a method for generating a library of different polynucleotide molecules, by ligating a double-stranded polynucleotide to a plurality of different target polynucleotide duplexes, the double-stranded polynucleotide comprising: (a) a first strand comprising an annealed portion and an overhang portion; and (b) a second strand consisting essentially of an annealed portion, wherein the second strand is complementary to and annealed to the annealed portion of the first strand.

The present invention relates to a method for generating a library of different polynucleotide molecules, by ligating a double-stranded polynucleotide to a plurality of different target polynucleotide duplexes, the double-stranded polynucleotide comprising: (a) a first strand comprising an annealed portion and an overhang portion; and (b) a second strand consisting essentially of an annealed portion, wherein the second strand is complementary to and annealed to the annealed portion of the first strand.

Aspects of the present disclosure relate to methods for modification and detection of methylated nucleotides. Embodiments are directed to detection of RNA methylation. Disclosed are methods and compositions for transcriptome-wide detection of N.sup.6-methyladenosine in mRNA. In some cases, methods for modifying a methylated nitrogenous base are described. Also disclosed are enzymes and other molecules useful for RNA methylation detection.

Aspects of the present disclosure relate to methods for modification and detection of methylated nucleotides. Embodiments are directed to detection of RNA methylation. Disclosed are methods and compositions for transcriptome-wide detection of N.sup.6-methyladenosine in mRNA. In some cases, methods for modifying a methylated nitrogenous base are described. Also disclosed are enzymes and other molecules useful for RNA methylation detection.

The present invention provides a fast and simple method for preparing a microbial profile of a human skin sample. The method can be performed using only portable devices, allowing for in-field profiling. The method is robust and has been found to work even with low DNA quantities and under difficult conditions.

The present invention provides a fast and simple method for preparing a microbial profile of a human skin sample. The method can be performed using only portable devices, allowing for in-field profiling. The method is robust and has been found to work even with low DNA quantities and under difficult conditions.

The present disclosure relates in some aspects to methods and compositions for analysis of a target nucleic acid, such as in situ detection of a region of interest in a polynucleotide in a tissue sample. In some embodiments, provided herein are templated ligation probes (e.g., RNA-templated ligation probes) and selector probes for generation of a circularized ligated probe comprising an insertion sequence of a selector probe, wherein the circularized ligated probe is amplified in a rolling circle amplification reaction to generate a product that is detected in the sample.