Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Specific primers for detecting mRNA expression level of bovine LPL (lipoprotein lipase) genes and fluorescent quantitative detecting kit

A technology of expression level and real-time fluorescence quantification, which is applied in the field of genetic engineering, can solve problems affecting the accuracy of quantification, and achieve the effects of good stability, simple and fast quantitative detection, and low experimental cost

Inactive Publication Date: 2015-09-09
LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since SYRB Green binds to all double-stranded DNA, false positives caused by primer-dimers, single-stranded secondary structures, and erroneous amplification products will affect the accuracy of quantification

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Specific primers for detecting mRNA expression level of bovine LPL (lipoprotein lipase) genes and fluorescent quantitative detecting kit
  • Specific primers for detecting mRNA expression level of bovine LPL (lipoprotein lipase) genes and fluorescent quantitative detecting kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The fluorescence quantitative PCR detection kit of LPL gene mRNA expression level of the present invention is made up of following components:

[0033] 2×SYBR GREEN MIX 5mL;

[0034] Positive control: 500 μL of standard LPL gene template;

[0035] Primer mixture 500μL;

[0036] Ultrapure water 5mL.

[0037] Among them, 2×SYBR GREEN MIX consists of the following components: Taq DNA polymerase 0.1 U / μL, dNTPs substrate 0.4 mmol / L, Mg 2+ 5.0 mmol / L, KCl 100 mmol / L, Tris·HCl 20 mmol / L at pH 8.3, 0.02% gelatin and SYBR Green dye.

[0038] Primer mixture: upstream primer 8 μmol / L, downstream primer 8 μmol / L. Wherein, the sequence of the upstream primer is: 5'-CCGCAGACAGGATTACAG-3',

[0039] The sequence of the downstream primer is: 5'-AGGAATGAGGTGGCAAGT-3'.

[0040] Standard LPL gene template: the concentration is 0.8 μmol / L, and the sequence of the LPL gene template is 5'-CCGCAGACAGGATTACAGGAGGAAAAGATTTTAGAGACATTGAAAGTAAATTTGCTCTCAGGACTCCCGAAGACACAGCTGAGGACACTTGCCACCTCA...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides specific primers for detecting the mRNA expression level of bovine LPL (lipoprotein lipase) genes. The nucleotide sequence of the forward primer is 5'-CCGCAGACAGGATTACAG-3'; and the nucleotide sequence of the reverse primer is 5'-AGGAATGAGGTGGCAAGT-3'. The invention further provides a corresponding detecting kit. The kit fulfills the purpose of simply and quickly detecting the transcription level of the LPL genes, and can be used for monitoring the mRNA expression state of different bovine species (including dairy cow, cattle and yak), different tissues and different periods. The kit has the advantages of high sensitivity, good stability and low experimental cost on the aspect of detecting the mRNA expression level of the genes. The kit can be used for monitoring the mRNA expression level of the LPL genes in the fat deposition process of different bovine species, interpreting the fat deposition principle and identifying the correlation between the genes and the animal fat deposition and meat quality.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a specific primer and a fluorescent quantitative PCR detection kit for detecting the expression level of bovine LPL gene mRNA. Background technique [0002] Polymerase chain reaction (Polymerase Chain Reaction, PCR) is the most commonly used technique in DNA manipulation technology in vitro. PCR technology puts template DNA, specific primers, dNTPs substrates, heat-resistant DNA polymerase, magnesium ions, etc. in the same buffer reaction system, and performs repeated thermal cycles of high-temperature denaturation, low-temperature annealing, and medium-temperature chain extension to achieve target DNA Fragments appear as 2 in the reaction solution n Double amplification (where n is the number of thermal cycles). [0003] Fluorescent quantitative PCR technology is based on ordinary PCR reaction, combined with real-time fluorescence detection technology a...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 裴杰褚敏郭宪包鹏甲梁春年丁学智阎萍冯瑞林王宏博朱新书
Owner LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com