Method and array for the replication and analysis of nucleic acids

摘要

The invention includes a method and array for replicating and analyzing one or a plurality of different target sequences in nucleic acid samples. The different reactions for replicating the target sequences occur simultaneously on an array with selectively heatable array elements with their reaction surfaces, some of which have been modified differently, and in the sample solution disposed thereabove. The target sequences are analyzed using molecular detection on reaction surfaces of said array that have been modified with probe strands. The target strands are replicated and analyzed simultaneously, successively, or alternately.

主权项

1. A method for replicating and electrochemically analyzing one or more different target sequences in nucleic acid samples, comprising:
a) at the same time, conducting different reactions on an array comprising selectively heatable array elements having temperature controlled reaction surfaces while the sample solution is in contact with the array elements, whereby the conducting of the different reactions effect the replication of one or more different target sequences; b) conducting electrochemical analysis of the one or more different target sequences on at least one of the individually temperature-controlled reaction surfaces, the surface being modified for the molecular detection of the one or more different target sequences by hybridization with probe sequences and being connected to an electrochemical measuring device for electrochemically detecting hybridization events as a working electrode thereof, the at least one of said reaction surfaces being modified with probe sequences for molecular detection of the target sequences using hybridization, wherein the temperature of said at least one modified reaction surface is individually controlled and the array element comprising the modified reaction surface is connected, as a working electrode, to an electrochemical measuring device for electrochemically detecting the hybridization events; c) replication of the one or more different target sequences being carried out in the following steps:
depositing the primer on the one or more different target sequences serving as templates;extending the primers by way of polymerase; andthermally decomposing nucleic acid double strands of the target sequences; wherein d) replication and electrochemical analysis are carried out in an order selected from: simultaneous with each other; first and second to each other, and in an alternating sequence with each other wherein the selectively heatable array elements having temperature controlled reaction surfaces include a first array element provided with an unmodified surface for facilitating a denaturing of a nucleic acid, the first array element heatable to greater than 90° C., a second array element provided with a surface modified with a polymerase layer for facilitating the extending of primer, the second array element heatable to a temperature of 20° C. to 70*C., and a third army element provided with a surface modified for hybridization with prob¢ strands that are bonded to the third array element by a thiol linker to gold.