Roche Diagnostics Gmbh

Boehringer Mannheim Gmbh

Heiko Wittor

Matthias Hinzpeter

Hermann Leying

Hans-Peter Fritton

Clemens Doppler

The invention describes a method of quantitatively detecting specific nucleotide sequences. Said method is essentially characterized in that a single-stranded nucleic acid, particularly mRNA, which has been isolated from a mixture, e.g. a biological sample, is hybridized in solution, with a polynucleotide sequence which is essentially complementary to the sequence to be determined; the nucleic acid is then immobilized on a solid phase, and the amount of bound hybrid is determined. It has proven to be particularly advantageous if the binding to the coated solid phase is accomplished with the aid of the specifically bindable chemical group which is coupled to the sequence to be determined or to the polynucleotide probe sequence via a linker.

Method of quantitatively detecting nucleic acids

Method and kit for the quantitative detection of specific oligonucleotide or polynucleotide sequences which is characterized in that a sample mixture containing RNA or single-stranded DNA is hybridized with an oligonucleotide or polynucleotide probe(s) which are complementary to the nucleotide sequence to be determined and carry(ies) a specificplly bindable and a detectable chemical group subsequently it is admixed with an agent that cleaves single-stranded polynucleotide sequences and the immobilized or non-immobilized nucleotide probe is determined after transfer into a suitable reaction vessel. It has proven to be particularly advantageous when a mixture is used which is composed of different cleaving reagents.

Method for the quantitative detection of specific nucleic acid sequences

The invention involves methods and kits useful in determining specific mRNA molecules. The methods can be carried in a single reaction vessel, in that both reverse transcriptase and amplification can be carried out in the same, heat stable reaction vessel.

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Tutzing, Germany

Heiko Wittor