SEQUENCE-SPECIFIC ANALYSIS OF NUCLEIC ACIDS

摘要

The invention relates to a method for the sequence-specific analysis of nucleic acids in a sample in which the nucleic acid is present at least partially as a double strand, which method uses electrochemical detection methods (e.g. with [OsO4(bipy)]). In particular, the methods comprise steps in which the at least partially double-stranded nucleic acid strands are converted by thermal denaturation to single strands which are termed target strands, and at least one nucleic acid strand designated protective strand is added, which protective strand can hybridize with a target strand, in order to form partial double-stranded segments, wherein the protective strands are shorter than the target strands, wherein the temperature of the sample is rapidly lowered to a temperature of below 5° C., preferably below 0° C. The invention also relates to devices which are suitable for these methods comprising a flow system in which the steps of the method can take place consecutively, having heatable sections for thermal denaturation as well as coolable sections for rapid cooling of the sample.

主权项

1. A method for the sequence-specific detection of nucleic acids in a sample, in which the nucleic acids are at least partially present as double strands, comprising
a) converting the at least partially double stranded nucleic acid strands into single strands termed target strands by thermal denaturation, b) adding at least one nucleic acid strand termed protective strand, which is able to hybridize with a target strand to form partially double stranded segments, wherein the protective strands are shorter than the target strands, wherein the temperature of the sample is rapidly lowered to a temperature of less than 5 ° C. c) labeling the remaining single stranded segments of the target strands via reaction with a redox marker, which reacts selectively with the double bond of the pyrimidine rings of the nucleic acid strands and allows an electroanalytically usable redox reaction on working electrodes, d) hybridizing the nucleic acid strands that are labeled in this manner on the surface of an electrode with probe strands that are immobilized thereon under replacement of the protective strands, and e) detecting the nucleic acid strands that are hybridized to the probe strands electroanalytically.