METHOD AND KIT FOR CHARACTERIZING RNA IN A COMPOSITION

• Main Authors: WEDLER ERIKA, WEDLER HOLGER
• Format: Patent
• Language: English
• Published: 13.11.2014
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; VINEGAR; WINE

The invention relates to a method for determining the sequence and/or quantity of a ribonucleic acid in a composition, comprising the steps of: i. providing a composition comprising one or more ribonucleic acids molecules (RNA), ii. hybridizing to said one or more RNAs, one or more two-part nucleic acid hybridization probes, wherein each probe comprises, a. a first nucleic acid molecule with a 3′-tail wherein said tail does not hybridize to an RNA in the composition, b. a second nucleic acid molecule with a 5′-tail wherein said tail does not hybridize to an RNA in the composition, c. wherein said first and said second nucleic acid molecules when and if hybridized to their target RNA lie on one single stranded RNA molecule separated from each other by between 2 and 1000 nucleotides, iii. covalently linking the hybridized 5′-tail of said first nucleic acid molecule to the hybridized 3′-tail of said second nucleic acid, wherein the linking is done by means of reverse transcription and subsequent ligation, iv. amplifying the linked molecules with primers that are specific for said first 3′-tail of said first nucleic acid molecule and said second 5′-tail of said second nucleic acid molecule and, v. sequencing the amplification products by means of next generation sequencing.