Lexogen Gmbh

Lukas Paul

Alexander Seitz

Torsten Reda

Irmlind Gabler

Petra Kubala

The present invention relates to methods for generating a labelled nucleic acid from an RNA comprising a 5' protecting group, said method comprises the steps of obtaining a mixture of template strands of nucleic acids, said mixture comprising said RNA and further potentially other nucleic acids without a 5&#x2032; protecting group, annealing at least one oligonucleotide primer to the template strand of said RNA and potentially other nucleic acids, and template sequence dependent extending said primer, thereby obtaining a complementary nucleic acid strand annealed to its template strand, or providing the RNA in duplex with a complementary nucleic acid strand annealed to its template strand, and optionally modifying the extension product of said nucleic acids without 5&#x2032; protecting group either on the 5&#x2032; end of the template strand or on the 3&#x2032; end of the complementary strand, or both, and labelling a complementary nucleic acid of a double stranded nucleic acid not modified, wherein therefore the labelled nucleic acid did have a 5&#x2032; protecting group on the RNA template strand, said 5&#x2032; protecting group being optionally removed after modification, and/or labelling a complementary nucleic acid of a double strand dependent on the presence of the 5&#x2032; protecting group on the complementary nucleic acids template RNA strand by double strand dependent ligation, thereby specifically generating a labelled nucleic acid complementary to an RNA at least originally comprising a 5&#x2032; protecting group.

5' protection dependent amplification

The present invention relates to the field of transcriptomics and provides a method for the controlled identification and/or quantification of transcript variants in samples, comprising providing a reference set of artificial polynucleic acid molecules simulating transcript variants and adding said reference set as external control to samples comprising transcript variants. The present invention further provides such a reference set, as well as a method to produce such a reference set.

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Vienna, Austria

Lukas Paul