Roche Diagnostics Operations, Inc.

Roche Molecular Systems, Inc.

Boehringer Ingelheim International Gmbh

Roche Diagnostics Gmbh

F. Hoffmann-la Roche Ag

Dirk Kessler

Dieter Heindl

Christoph Seidel

Angelika Roesler

Wilma Thuer

Hans-Peter Josel

Frank Bergmann

Nancy J Schoenbrunner

Matthew Russell Netherton

Alison Tsan

Stefanie Froehner

Lars Van Der Veen

Christian Weilke

Gunter Lampert

Siegfried Schneider

Ralph Brueckner

Oliver Kraemer

Darryl Mcconnell

Davide Gianni

Andreas Steffen

Christian Smethurst

Dorothea Rudolph

Michael Sanderson

Anke Baum

Fabio Savarese

Laetitia Janine Martin

Michael Gmachl

Marco Hofmann

Harald Weinstabl

Christiane Kofink

Tobias Wunberg

Juergen Ramharter

Andreas Mantoulidis

Andreas Gollner

Harald Engelhardt

Ulrich Reiser

Nancy Schoenbrunner

The present invention includes a method of allele-specific amplification, utilizing an allele-specific oligonucleotide, at least partially complementary to more than one variant of the target sequence, but having at least one selective nucleotide complementary to only one variant of the target sequence and incorporating both a nucleotide with a base covalently modified at the exocyclic amino group and a modified phosphate.

Primers with modified phosphate and base in allele-specific PCR

The trivalent phosphorous atom of a compound is reacted with a reagent in such a manner that a stable phosphate mimetic or a specifier is formed. Phosphoramidites with a phosphorous atom containing at least one hydroxyl residue which is provided with a protective group are reacted for this purpose with a free hydroxyl group: In the first synthesis cycle the hydroxyl group is linked to a solid support via a cleavable or non-cleavable linker. In further synthesis cycles the hydroxyl group is created by cleavage of the protective group from the growing oligomer. This results in formation of a phosphorous acid triester which is reacted with azides. By selecting suitable monomers for the synthesis which have a defined stereoconformation compounds of Formula 1 are produced in a stereocontrolled manner. 

Directed synthesis of oligophosphoramidate stereoisomers

The current invention restates substituted indole nucleosides as both terminal as well as internal building blocks of labeled oligonucleotide probes for the detection, analysis and quantitation of nucleic acids. The substituent comprises a linker and a detectable group or a linker and a reactive group for post synthesis coupling. These modified nucleosides grant access to a wide application area. These new substituted indole nucleosides can be used as labeling reagents for the facile preparation of e.g. optimized hybridization probes, Taqman-probes, or molecular beacon probes.

Labeling reagent

The present invention concerns modified oligonucleotides and processes for their production wherein these oligonucleotides contain at least once the structure P&#x2550;N-Acc where Acc is an electron acceptor or an electron acceptor substituted with a residue R and R is any organic substituent.

Polynucleotide containing a phosphate mimetic

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Peiting, Germany

Dirk Kessler