Method for detecting target DNA sequence, gene chip adopting same and application of gene chip

Abstract

The invention discloses a method for detecting a target DNA sequence. The method comprises the following steps: 1) an RNA probe contacts with a target DNA to be annealed, wherein one end of the RNA probe is fixed on a substrate, the RNA probe contains a detection sequence close to the fixed end and a marking sequence close to the free end and the target DNA has a segment which is complimentary tothe detection sequence in the RNA probe; 2) ribonuclease is used to hydrolyze the obtained DNA after annealling, wherein the RNA component in the RNA heteroduplex releases the target DNA and the marking sequence of the RNA probe; and 3) the free target DNA and marking sequence are removed through washing, the marking DNA with a marker is bound with the RNA probe of the detection sequence to perform sequence analysis on the target DNA, wherein the RNA probe is not hydrolyzed and is fixed on the substrate, and the marking DNA is complimentary to the marking sequence of the RNA probe. By adopting the method to detect the target DNA, the target gene does not require PCR amplification and the target DNA is directly detected, thus avoiding adopting the false negative and false positive results of the PCR amplification and increasing the detection accuracy rate.

Description

technical field [0001] The invention relates to a molecular biological detection method of a gene sequence, a gene chip for detecting a gene sequence using the method, and an application of the method. Background technique [0002] At present, the detection of genetic background-related genes (disease-causing genes, abnormal tumor genes), bacteria, viruses, and transgenic components of transgenic animals and plants is mainly through the design of primers and PCR amplification of the corresponding DNA fragments, and then through the subsequent detection platform. achieve the purpose of testing. Detection by PCR amplification can easily lead to false negative (PCR amplification failure) or false positive (PCR amplification contamination) and other results, thereby affecting the correctness and efficiency of gene sequence detection. In addition, the amount of target genes required for PCR amplification is large, and it is difficult to meet the detection requirements in the cas...

AI Technical Summary

Problems solved by technology

Detection by PCR amplification can easily lead to false negative (PCR amplification failure) or false positive (PCR amplification contamination) and other results, thus affecting the accuracy and efficiency of gene sequence detection

In addition, the amount of target gene required for PCR amplification is large, and it is difficult to meet the detection requirements in the case of scarce samples.

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