Method for low-sequence-dependence and high-order constant-temperature-index amplification testing of microRNA

A constant temperature exponential amplification and sequence technology, which is applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems such as sequence dependence, and achieve low sequence dependence, good specificity, and simple amplification system Effect
CN109136353AActive Publication Date: 2019-01-04SUN YAT SEN UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
SUN YAT SEN UNIV
Publication Date
2019-01-04

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Abstract

The invention discloses a method for low-sequence-dependence and high-order constant-temperature-index amplification testing of microRNA. The method comprises the steps of designing specific templatesT1 and T2 which can detect target microRNA; mixing the templates T1 and T2, DNA polymerase, incising incision enzyme and dNTP; reacting at a temperature of 35-50 DEG C; detecting an amplification curve; and judging whether a sample contains the target microRNA according to the amplification curve and the standard curve. By the method, the specific content of the target microRNA in the sample canbe calculated further. The method is high in amplification efficiency and high in sensitivity, and the analyzing time is shortened; the method has low sequence dependence on the target microRNA, and the universality is high; a detection linear range is wide, and the specificity is good; and an amplification system is simple, can be developed into a kit and is popularized to markets.
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Description

technical field

[0001] The invention belongs to the field of nucleic acid quantitative detection, and in particular relates to a method for detecting microRNA (hereinafter referred to as miRNA) by low sequence dependence and high-order constant temperature exponential amplification. Background technique

[0002] miRNA is a type of endogenous non-coding small molecule RNA, about 18-24 nucleotides in length, it has a regulatory effect on gene expression, and actively participates in cell growth, differentiation and apoptosis in vivo. A large number of research results show that the expression level of miRNA is closely related to cancer or certain diseases. Therefore, the quantitative detection of miRNA not only helps to understand its mechanism of action, but also has great significance for the diagnosis and treatment of diseases and the development of related gene medicines. However, due to the short sequence of miRNAs and high sequence similarity, the difference between hom...

Claims

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