Detection process for synthesizing microfluid chip by in-situ method

Abstract

The invention discloses a detection method synthesizing microfluidic chips in situ, which comprises the following steps that: 1) a sequence of non-coding small RNAs reported on the prior model plant is utilized, sequence information of miRNA is preferably selected, and complementary oligonucleotide hybridization probes are designed according to the sequence of a sense strand or an antisense strand of miRNA; 2) an oligonucleotide chip is prepared by an in-site synthetic method; 3) small RNAs smaller than a 50 oligonucleotide group are extracted from a plant to be detected, then are hybridized with the probes in the chip after making end labeling, and the result of chip hybridization is obtained to perform qualitative and quantitative analysis. The detection method can be used for the validation of miRNA (and other sNC-RNAs) obtained through the forecast by a bioinformatics method, can also be used for the detection and identification of miRNA (and other sNC-RNAs) in new plant species, and can also be used for the determination of miRNA (and other sNC-RNAs) variation relation under different treatment conditions (eg. under the condition of virus infection).

Description

A detection method using in situ synthesis of microfluidic chips technical field The invention relates to a method for detecting and identifying small non-coding RNAs in plants, in particular to a method for detecting known miRNAs using a chip. Background technique MicroRNA (miRNA) is a class of non-coding small RNAs widely present in eukaryotes. They regulate post-transcriptionally by complementing target mRNA sequences, and play an important role in the regulation of gene expression in plant growth and development. Conservation is an indicator of the function of miRNAs, and conserved miRNAs play very important roles in the regulation of conserved genes, such as leaf and flower morphology and signal transduction. Unconserved miRNAs may play more specific roles in specific plant species, such as cotton fiber differentiation and elongation. miRNAs are highly conserved in different populations of animals or plants. For example, in animals miRNAs such as miR7 and miR18 are ...

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Problems solved by technology

However, the above studies are limited to high-throughput detection of miRNAs that have been reported in one species, and so far there has been no report of using known miRNAs of existing species to detect and discover unknown miRNAs of other species

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