Method for analyzing rna
AssigneeCureVac AG
InventorsAniela WOCHNER, Tilmann ROOS, Fabian Johannes EBER, Philipp HOFMANN
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Abstract
The present invention relates to the field of RNA analysis. In particular, the invention concerns the use of one or more nucleic acid molecules for the analysis of an RNA molecule. In particular, the method is suitable for use in quality control during or following production of RNA. Furthermore, the present invention provides methods for analyzing a mixture of RNA molecules or an RNA population.
Claims93
A method for analyzing a sample comprising in vitro transcribed RNA, comprising determining the presence, integrity and/or quantity of at least one RNA species having a sequence comprising a target sequence in said sample using a nuclease protection assay or a molecular beacon assay.
A method for analyzing a sample comprising in vitro transcribed RNA, comprising determining the presence, integrity and/or quantity of at least one RNA species having a sequence comprising a target sequence in said sample using reverse transcription followed by quantitative PCR (RT-qPCR), wherein said analysis is independent of the target sequence of the at least one RNA species.
Method according to claim 32, wherein the RNA species further comprises a sequence located 5′ and/or 3′ of the target sequence and wherein the at least one set of PCR primers binds to said sequence located 5′ and/or 3′ of the target sequence.
Method according to any one of the preceding claims, wherein said sample comprising in vitro transcribed RNA comprises different RNA species.
Method according to any one of the preceding claims, wherein the analysis comprises determining the presence, integrity and quantity of the at least one RNA species in said sample comprising in vitro transcribed RNA.
Method of analyzing a sample comprising different RNA species, each having a sequence comprising a target sequence and being prepared by RNA in vitro transcription, the method comprising determining the presence, integrity and/or quantity of each of the RNA species present in said sample using a nuclease protection assay, RT-qPCR or a molecular beacon assay.
Expression vector comprising in 5′ to 3′ direction a promoter and operably linked thereto a nucleic acid sequence encoding a target sequence and at least one synthetic sequence located 5′ and/or 3′ of the target sequence.
Expression vector comprising in 5′ to 3′ direction a promoter and operably linked thereto a nucleic acid sequence encoding a 5′ untranslated region, a multiple cloning site and a nucleic acid sequence encoding a 3′ untranslated region, wherein the 5′ and/or 3′ untranslated region comprises a synthetic sequence.
Nucleic acid molecule consisting of the sequence according to any one of SEQ ID Nos. 5 to 8, wherein a fluorophore is attached to one end of the nucleic acid molecule and a quencher is attached to the other end of the nucleic acid molecule.
Dependencies are inferred from the claim text.