Roche Diagnostics Gmbh

Boehringer Mannheim Gmbh

Roche Diagnostics Operations, Inc.

Roche Sequencing Solutions, Inc.

F. Hoffmann-La Roche Ag

Bruno Frey

Harald Sobek

Klaus Kaluza

Michael Jarsch

Dieter Heindl

Frank Bergmann

Waltraud Ankenbauer

Manfred Schmidt

Mosè Rossi

Garabed Antranikian

Francesca Maria Pisani

Britta Villbrandt

Dietmar Schomburg

Rainer Mueller

Christoph Kessler

Johann-Peter Thalhofer

Rainer Schmuck

Johannes Auer

Bernhard Suppmann

Gudrun Schmitz-Agheguian

Max Lechner

Markus Pajatsch

Boehlke Kristina

Peter Pallua

Kristina Boehike

Barbara Prinz

Provided herein is a new class of nucleic acid tagging molecules which are essentially free of homopolymer stretches. The tagging molecules can tag a plurality of individual target molecules for detection with a high degree of accuracy. The tagging molecules can be used to tag at least 10or 10individual target molecules. The tagged individual target molecules can be subjected to high throughput sequence analysis.

Sequence tags

Provided herein is a new class of nucleic acid tagging molecules which are essentially free of homopolymer stretches. The tagging molecules can tag a plurality of individual target molecules for detection with a high degree of accuracy. The tagging molecules can be used to tag at least 105 or 106 individual target molecules. The tagged individual target molecules can be subjected to high throughput sequence analysis.

The present invention relates to thermostable mutants of B-type DNA polymerases comprising a Y-GG/A amino acid motif between the N-terminal 3'-5&#x2032;-exonuclease domain and the C-terminal polymerase domain whereas the tyrosine of the Y-GG/A amino acid motif is mutated and whereas these mutant DNA polymerases are suitable for PCR.

Mutant B-type DNA polymerases exhibiting improved performance in PCR

The invention concerns polymerase chimeras which are composed of amino acid fragments representing domains and which combine properties of naturally occurring polymerases that are advantageous with regard to a particular application. It has surprisingly turned out that the domains from the various enzymes are active in the chimeras and exhibit cooperative behavior. In addition the present invention concerns a process for the production of the chimeras according to the invention and the use of these chimeras for the synthesis of nucleic acids e.g. during a polymerase chain reaction. Moreover the present invention concerns a kit which contains the polymerase chimeras according to the invention.

Polymerase chimeras

The heterologous expression of the reverse transcriptase from the Avian Myeloblastosis Virus (AMV-RT) in prokaryotic cells and in particular() is described in the present invention. The invention also includes certain measures to simplify the purification of the heterodimeric AMV-RT.

Method for producing an active heterodimeric AMV-RT in prokaryotic cells

Thermolabile enzyme with uracil-DNA-glycosylase activity which is in particular characterized by a high degree of purity, short half-lives and a content of contaminating foreign activities of less than 2%, a process for its isolation as well as the use thereof to remove the base uracil from DNA and in particular from PCR products containing uracil. The enzyme is obtainable from gram-positive microorganisms such as e.g. Arthrobacter or Micrococcus.

Thermolabile uracil-DNA-glycosylas, process for its preparation and use for removing uracil from DNA

The new type II restriction endonuclease SexAI has the following recognition sequence and preferably cleaves at the cleavage site defined by the mark: ##STR1## It is preferably obtainable from microorganisms of the genus Streptomyces.

Type II restriction endonuclease SexAI

The new type II restriction endonuclease Ssp4800I has the following recognition sequence: ##STR1## and preferably cleaves at the cleavage site defined by the arrows. It is preferably obtainable from microorganisms of the genus Streptomyces.

Type II restriction endonuclease Ssp4800I

The novel type II restriction endonuclease SwaI has the following recognition sequence: ##STR1## and preferably cleaves at the cleavage site indicated by the line. It is preferably obtainable from microorganisms of the genus Staphylococcus.

Type II restriction endonuclease SwaI

The type III restriction endonuclease SgrAI has the following recognition sequence: ##STR1## cleaves DNA at the cleavage site indicated by the arrows, and is preferably obtainable from microorganisms of the genus Streptomyces. It can be used to recognize and cleave the double-stranded DNA sequence ##STR2## and its complementary sequence.

Type II restriction endonuclease SgrAI

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Penzberg, Germany

Bruno Frey