Nucleic acid detection method based on liquid chip technology

Abstract

The invention discloses a nucleic acid detection method based on a liquid chip technology. The nucleic acid detection method comprises the following steps: 1) carrying out PCR amplification on a nucleic acid sequence to be detected; 2) preparing a liquid chip; 3) carrying out nucleic acid hybridization; and 5) analyzing a result. A forward primer used in the step (1) is a common primer with a section of specific binding sequence, and the specific binding sequence in the forward primer is defined as an Anti-Probe sequence; and the quantity of the forward primer is equal to that of a reverse primer. The obtained PCR amplification product is a double-stranded DNA product, the double-stranded DNA product is provided with a marker, and the Anti-Probe sequence exists at one end of the double-stranded DNA product in a free single-stranded state. The multi-target detection is realized by adopting multiplex-PCR, and the enrichment of a plurality of nucleic acid target sequences is realized by applying the specific primers. After a specific Anti-Probe binding sequence at the 5'end of an enriched product is combined with Probe which is connected to encoding microspheres and can be complementary with the Anti-Probe binding sequence, the classified detection of a plurality of check table sequences is realized.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a nucleic acid detection method based on liquid chip technology. Background technique [0002] Nucleic acid detection generally requires two processes of PCR amplification and nucleic acid hybridization of the nucleic acid to be tested, and then the product is analyzed. Among them, PCR (Polymerase Chain Reaction) is a molecular biology technique used to replicate and amplify specific DNA fragments in vitro; the principle of PCR is: DNA is denatured to form a single strand at 95°C in vitro; Primers combine with single strands according to the principle of complementary base pairing; DNA polymerase synthesizes complementary strands along the direction from phosphoric acid to five-carbon sugar (5'-3') at ~72°C, so that the product obtained by PCR is usually double-stranded DNA . Nucleic acid hybridization is a process in which complementary nucleotide sequences (DNA and ...

AI Technical Summary

Problems solved by technology

However, for different nucleic acids to be tested, it is still necessary to design and synthesize probes that match the nucleic acids to be tested. The project is still time-consuming and the research and development costs are still high.

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