Signal detection method for gene chip of oligonucleotide with length more than 50 basic groups

Abstract

A method for detecting the signals of gene chip whose oligonucleotide has a length more than 50 bases includes such steps as PCR amplification of target gene, detecting, boiling positive product for modifying it, cooling on ice, wetting the gene chip by phosphoric acid solution, prehybridizing in prehybridizing liquid, adding said modified DNA, hybridizing, water with different lotions, dyeing, rinsing, and detecting signal under UV light or fluorescent microscope.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a signal detection method of a gene chip of an oligonucleotide with a length greater than 50 bases. Background technique [0002] Since Adams proposed expressed sequence tag (expressed sequence tag) in 1992 and the subsequent establishment of large-scale expressed sequence tag technology, as well as the development of biotechnology and the needs of life science development, gene chip technology has emerged as the times require. The essence of gene chips (Biochips) is to arrange a series of addressable biorecognition molecules fixed at a certain position in an orderly array on a small substrate (R.J. Lipshutz et al., Bio Techniques 19 (1995), 442 -447; S.P.Fodor et al., Nature 364(1993), 555-556; S.P.Fodor et al., Science 251(1991), 767-773; A.C.Pease et al., Proc.Natl.Acad.Sci.USA 91 (1994), 5022-5026). Since the original gene chip was mainly used for DNA sequence deter...

AI Technical Summary

Problems solved by technology

At present, signal detection methods for gene chips mainly include fluorescent signal detection methods labeled with Cy5 / Cy3 and isotope-labeled signal detection methods, and these methods are currently mostly used in signal detection of high-throughput gene chips, but regardless of the application The signal detection methods all need to label the probe, and the application of Cy5 / Cy3 fluorescent labeling requires Cy5 and Cy3 to be carried out twice respectively. Using isotope labeling can not only cause personal injury to the operator, but also cause nuclear radiation pollution to the environment.

Images