Primer, kit and detection method for detecting 245bp deletion alternative spliceosome of LEPR gene

Abstract

The invention relates to a primer, a kit comprising the primer and a detection method for detecting a 245bp deletion alternative spliceosome of an LEPR gene and aims at solving the problems of tedious program, long consumed time and low accuracy. The detection method comprises the following steps: with cDNA as a tempelate and a primer pair ACTB as a primer, and amplifying an ACTB reference gene by virtue of real-time fluorescence quantitative PCR; with a primer pair P as a primer, carrying out real-time fluorescence quantitative PCR amplification, wherein the reaction system of the real-time fluorescence quantitative PCR amplification consists of 2*SYBR@Premix Ex TaqTMII, ultrapure water with RNase removed, a primer P-F or ACTB-F, a primer P-R or ACTB-R, and the cDNA tempelate, and the reaction method comprises the following steps: initial denaturation, denaturation, annealing and extension; and analyzing a qPCR real-time monitoring result, calculating by using 2-delta CT relative quantification, and carrying out difference significance test by SPSS version 20.0. The primer, the kit and the detection method are accurate, rapid and convenient and strong in applicability.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting LEPR A primer for a gene 245bp deletion alternative splice body, and a kit and detection method comprising the primer. Background technique [0002] Leptin is mainly a small peptide hormone secreted by the white adipose tissue of non-avian vertebrates, which binds to the leptin receptor (LEPR) to exert its biological function. However, functional studies of LEPR are limited due to the lack of avian endogenous Leptin. Recently, birds Leptin The successful cloning of the gene has pushed the research of bird Leptin and LEPR to a new climax. birds LEPR The widespread expression characteristics of gene organization have been proved, but the identification and isolation of multiple alternative splicing forms are relatively slow, which will hinder the LEPR The study of the physiological function of genes. Therefore, it is of great significance to use molecular ...

AI Technical Summary

Problems solved by technology

[0004] In view of the above situation, in order to overcome the defects of the prior art, the purpose of the present invention is to provide a method for detecting the alternative splicing of LEPR gene 245bp deletion The primers for the body and the kits and detection methods containing the primers can effectively solve the problems of cumbersome procedures, long time-consuming and low accuracy of the existing detection methods

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