Agct Gmbh

Norbert Basler

Dmitry Cherkasov

Claus Becker

Andreas Mueller-Hermann

Hans-Joerg Hess

Christian Grunwald

Andreas Muller-Hermann

Petra Van Husen

Hans-Jörg Hess

Andreas MÜLLER-Hermann

A method for amplification of nucleic acids in which substantially use is made of the fact that a pre-defined nucleic acid chain (target sequence) can be multiplied/amplified in the presence of a target sequence-specific activator oligonucleotide. The target sequence-specific activator oligonucleotide causes the separation of re-synthesized complementary primer extension products by strand displacement, so that a new primer oligonucleotide can attach to the respective template strand. The thus formed complex of a primer oligonucleotide and a template strand can initiate a new primer extension reaction. The thus formed primer extension products in turn function as templates, so that an exponential amplification reaction results. Amplification of a particular target sequence takes place more efficiently in case of perfect match complementary base pair formation between the activator oligonucleotide and the corresponding target sequence. Mismatches between the activator oligonucleotide and a particular target sequence can result in less efficient amplification. The efficiency of synthesis of perfect match target sequences and mismatch sequences can be measured and compared.

Process for the enzymatic synthesis and amplification of nucleic acids

It is disclosed a method for the detection of an amplification of nucleic acids in which substantially use is made of the fact that a pre-defined nucleic acid chain (target sequence) can be multiplied/amplified in the presence of a target sequence-specific activator oligonucleotide. The target sequence-specific activator oligonucleotide causes the separation of re-synthesized complementary primer extension products by means of strand displacement, so that a new primer oligonucleotide can attach to the respective template strand. The thus formed complex of a primer oligonucleotide and a template strand can initiate a new primer extension reaction. The thus formed primer extension products in turn function as templates, so that an exponential amplification reaction results. Amplification is detected by a detection system.

Method for indicating the progress of amplification of nucleic acids and kit for performing the same

A method for amplification of nucleic acids in which substantially use is made of the fact that a pre-defined nucleic acid chain (target sequence) can be multiplied/amplified in the presence of a target sequence-specific activator oligonucleotide. The target sequence-specific activator oligonucleotide causes the separation of re-synthesized complementary primer extension products by strand displacement, so that a new primer oligonucleotide can attach to the respective template strand. The thus formed complex of a primer oligonucleotide and a template strand can initiate a new primer extension reaction. The thus formed primer extension products in turn function as templates, so that an exponential amplification reaction results.

The invention relates to a novel method for the enzymatic marking of nucleic acid chains (target sequences) by means of nucleotide conjugates. Said nucleotide conjugates are able, under reaction conditions, to specifically bind to the target sequence and integrate into the complementary growing strand by means of a polymerase. The nucleic acid chains marked by such conjugates can be bound to the solid phase. The marking can be carried out parallel to the enzymatic amplification of target sequences.

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Grosshansdorf, Germany

Norbert Basler