Method for detecting miRNA (micro ribonucleic acid) in stomach juice

A detection method and gastric juice technology, applied in the field of miRNA detection in gastric juice, can solve the problems of low miRNA, difficult miRNA extraction and detection, no specific extraction miRNA detection method, etc., and achieve the effect of convenient centrifugal separation
CN102127601AInactive Publication Date: 2011-07-20NINGBO UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
NINGBO UNIV
Publication Date
2011-07-20
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a method for detecting miRNA (micro ribonucleic acid) in stomach juice, which comprises stomach juice extraction and neutralization, RNA release, chloroform extraction, isopropanol precipitation, ethanol washing, RNA reverse transcription, PCR (Polymerase Chain Reaction) detection and other steps. In the detection method, a sodium hydroxide solution is used to liquefy viscous components in the stomach juice, so that mucoproteins and polysaccharides are suspended in the solution, and the stomach juice is neutralized; trypsinase is added to carry out digestion, so that the mucoproteins and other proteins can be hydrolyzed, and cell components in the stomach juice are loosened at the same time, thereby facilitating centrifugal separation; and a Trizol solution is added to carry out repeated blow and beat, so that the RNA is completely released and separated from the polysaccharides, the mucoproteins and other impurities, thereby solving the problem that the RNA extraction can be disturbed by the acidity of the stomach juice as well as polysaccharides, mucoproteins and the like in the stomach juice. Thus, the invention can eliminate the disturbance caused by polysaccharides, proteins and other impurities in the stomach juice, complete the efficient extraction of miRNA in the stomach juice and carry out fluorescent quantitative PCR detection on the extracted miRNA.
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Description

Technical field

[0001] The invention relates to a detection method of miRNA, in particular to a detection method of miRNA in gastric juice. Background technique

[0002] RNA in organisms can be divided into two categories: coding RNA and non-coding RNA. Non-coding RNA can be divided into long-chain non-coding RNA and short-chain non-coding RNA; the latter is more important including microRNA (microRNA, miRNA), which is 19-24 nucleotides in length. In the normal development of organisms, miRNAs play a pivotal role. They play an important negative regulatory role in gene expression through the cleavage or translation inhibition of target mRNA, and have important functions such as affecting cell growth, differentiation and apoptosis. In the physiological abnormal variation of different tissues and cells, some miRNAs have obvious abnormal expression.

[0003] The analysis of miRNA requires small RNA extraction and detection. For small RNA extraction, such as Chinese patent applicatio...

Claims

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