Primer and kit for detecting 127bp deletion variable spliceosome of LEPR gene and detecting method

Abstract

The invention discloses a primer for detecting 127bp deletion variable spliceosome of LEPR gene and a kit containing the same and a detecting method. The problems of tedious procedure, long time consumption and low accuracy can be effectively solved. The detecting method comprises the following steps: by taking a cDNA to be tested as a template and taking a primer pair ACTB as a primer, carrying out real-time fluorescence quantification PCR amplification on an ACTB reference gene; and by taking a primer pair P as a primer, carrying out real-time fluorescence quantification PCR amplification on 127bp deletion alternative spliceosome of LEPR gene, wherein the reaction system of the fluorescence quantitative PCR amplification includes ultrapure water for removing the RNA enzyme, a primer P-F or ACTB-F, a primer P-R or ACTB-R and a cDNA template, and the reaction method comprises the steps of pre-denaturing, denaturing, annealing, extending and circulating for 35-45 times, analyzing the qPCR real-time monitoring result, calculating by 2-Delta CT relative quantification method and performing SPSS version 20.0 significance test of difference. The method is accurate, rapid, 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 the 127bp deletion alternative splice body of the gene, a kit containing the primer and a detection method. 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 mol...

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 alternative splicing of LEPR gene 127bp 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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