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RNA detecting kit and method

A kit and oligonucleotide technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of increased total reaction time, increased reverse transcription reaction time, and increased Ct value, etc.

Pending Publication Date: 2019-04-26
HEIMBIOTEK INC
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Problems solved by technology

However, in the case of this conventional detection method, due to its long length, the diffusion of the reverse transcription primer in the reaction solution is slow, and thus the reverse transcription reaction time increases, resulting in an increase in the total reaction time
Therefore, the problem with conventional detection methods is that the C in real-time PCR t The value becomes larger and the reactivity decreases

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[0024] Definition of terms

[0025] As used herein, the term "oligonucleotide" generally refers to a polymer of nucleotide monomer units, which are short single-stranded nucleic acid molecules composed of 13 to 25 nucleotides. In some instances, the term may refer to nucleic acid molecules composed of fewer than 13 nucleotides, including 6-mers, 7-mers, 8-mers, 9-mers, 10-mers, 11-mers, and 12-mers , or nucleic acid molecules consisting of more than 25 nucleotides. As used herein, the term "polynucleotide" is generally a polymer of nucleotide units longer than the oligonucleotides defined above, but is also used interchangeably with the term "oligonucleotide". Polynucleotides include both single- and double-stranded nucleic acid molecules.

[0026] As used herein, the term "RNA molecule" refers to a multimeric molecule that plays multiple roles in the coding, decoding, regulation, and expression of genes, and is generally distinguished from double-stranded DNA in that it fo...

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Abstract

The present invention relates to a kit and a method for detecting an RNA molecule, particularly, a small-size RNA molecule such as miRNA. The present invention provides an RNA detecting kit and methodin which short primers for reverse transcription are used in a reverse transcription process with the aim of rapidly diffusing the primers in a reverse-transcription reaction solution and thus reducing the reverse-transcription reaction time, enabling the overall PCR reaction rate to be increased and RNA to be detected stably and rapidly. In addition, the present invention provides an RNA detecting kit and method in which, after a reverse transcription process, extension is performed on a single-stranded cDNA by use of a primer for extension, far longer than a primer for reverse transcription, and thus a template long enough to conduct PCR amplification can be obtained.

Description

technical field [0001] The present invention relates to kits and methods for detecting nucleic acids, more particularly to kits and methods for detecting RNA. Background technique [0002] MicroRNA (MicroRNA) (hereinafter abbreviated as "miRNA") is a non-coding RNA consisting of about 22 ribonucleotides. miRNAs are known to regulate RNA silencing at the transcriptional stage and gene expression at the post-transcriptional stage. This function of miRNAs is carried out by base pairing with complementary sequences in their target mRNAs. Since miRNAs were first discovered in Caenorhabditis elegans (C. elegans), various miRNAs have been reported one after another and are known to regulate gene expression in animals, plants and viruses. In addition, it is known that dysfunction caused by mutations in miRNA genes may contribute to certain diseases, such as cancer. Therefore, the importance of miRNAs has been increasingly recognized. [0003] Since miRNAs are known to be very sh...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6827
CPCC12Q1/6853C12Q1/6848C12Q2521/107C12Q2525/155C12Q2525/161C12Q2525/173C12Q2525/191C12Q2525/207C12Q2531/113C12Q2533/10C12Q2561/113
Inventor 李哉勋
Owner HEIMBIOTEK INC
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