Nucleic acie detecting method using bar code-type gene chip

Abstract

A preparation process of bar code-type gene chip is to fix oligonucleotide segments with specific base sequence onto the surface of substrate in microarray mode. The forward primer upstream sequence with mononucleotide site specificity to be detected is made to complement and pair with bar code, the middle is the limited enzyme incising site, and the downstream sequence is made to pair with the target sequence. There is a biotin or a fluorescent group mark in the 5'-end, and a nucleotide incapable of pairing with target sequence is introduced to the place of 1-4 nucleotides apart from the 3'-end upstream through PCR, purification, enzyme incising and hybrid elution with the bar code gene chip. The bar code base sequence has composition independent on target nucleic acid to be detected and has detected segment length within 30 nucleotides, and this reduces the cross hybridizing and error hybridizing.

Description

Technical field: [0001] The invention relates to a method for detecting nucleic acid to be detected by using a gene chip, in particular to a method for detecting nucleic acid to be detected by using a barcode gene chip, which can be used for nucleic acid sequence analysis and belongs to the technical field of biological information engineering. Background technique: [0002] Gene chip technology is to immobilize gene probes on the surface of solid phase carrier in the form of microarray, and analyze the nucleic acid species and content of the sample solution to be tested according to the principle of nucleic acid base pairing. Most of the types of gene chips at present are that nucleic acid fragments are fixed on the surface of glass slides (Patricia L.Dolan, Yang Wu, Linnea K.Ista, Robert L.Metzenberg) of activation treatment (amination, formylation, isothiocyanation) , Mary Anne Nelson, and Gabriel P. Lopez. Robust and efficient synthetic method for forming DNA microarrays...

AI Technical Summary

Problems solved by technology

In the current detection, the nucleic acid length of the detection sample is too long, all of which are more than 100 nucleotides in length. Cross-hybridization and mis-hybridization are very common, and the accuracy rate is only 60%-70%.

In addition, because the nucleic acid to be tested must be labeled with a fluorescent group for analysis, the difficulty of operation is increased.

When the sample to be tested contains a variety of nucleic acid samples and needs to be analyzed, the difference between the sample markers in a reaction solution makes the data untrue, etc., which limits the popularization and application of the gene chip