Primer for detecting mango EIF1A (eukaryotic translation initiation factor) gene and real-time fluorescent quantitative PCR (polymerase chain reaction) method

A fluorescent quantitative and genetic technology, applied in the field of fluorescent quantitative PCR detection, can solve the problem of undiscovered EIF1A gene distribution and achieve the effect of improving the amplification efficiency

Inactive Publication Date: 2018-04-20
GUANGXI UNIV +1
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, there is no research on the distribution of mango EIF1A gene in different mango tissues.

Method used

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  • Primer for detecting mango EIF1A (eukaryotic translation initiation factor) gene and real-time fluorescent quantitative PCR (polymerase chain reaction) method
  • Primer for detecting mango EIF1A (eukaryotic translation initiation factor) gene and real-time fluorescent quantitative PCR (polymerase chain reaction) method
  • Primer for detecting mango EIF1A (eukaryotic translation initiation factor) gene and real-time fluorescent quantitative PCR (polymerase chain reaction) method

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Experimental program
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Effect test

Embodiment

[0037] 1. Experimental materials:

[0038] Different tissue samples of Four Seasons mango: including leaves, stems (lignified), flowers, fruits, sampled at different periods respectively, after quick freezing, and stored at -80°C (this example only gives an optimal implementation temperature , the inventor found through research that when the storage temperature is -70°C to -80°C, it can meet the temperature requirements for the storage of the samples of this application) refrigerator.

[0039] 2. Total RNA extraction of the target gene:

[0040] 2.1 Extraction of total RNA from mango leaves:

[0041] (1) Get old mango leaves, grind 100 mg of mango old leaves into powder with nitrogen liquid, transfer to 2.0 mL centrifuge tube I, and then add 1 mL of cell lysate;

[0042] (2) Add 300 μL of deproteinized solution to the centrifuge tube in step (1), mix for 30 seconds, take the lower layer solution and transfer it to centrifuge tube II containing 200 μL chloroform, fully mix t...

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Abstract

The invention relates to the technical field of a fluorescent quantitative PCR (polymerase chain reaction) detection method, in particular to a primer for detecting a mango EIF1A (eukaryotic translation initiation factor) gene and a real-time fluorescent quantitative PCR method. The method uses the fluorescent quantitative PCR technology for analyzing the EIFIA gene expression quantity of different tissues of the mango; the MiGAPDH is used as a reference gene; the reference gene primer and a target gene primer are designed; reverse transcription cDNA is used as a template for detecting the expression level of the target gene on Light 480. The detection result is subjected to statistical analysis; the expression and distribution condition of the EIF1A gene in different tissues of the mangocan be fast and accurately detected; the scientific basis is provided for cloning and utilizing the gene.

Description

【Technical field】 [0001] The invention relates to the technical field of fluorescent quantitative PCR detection methods, in particular to a primer for detecting mango EIF1A gene and a real-time fluorescent quantitative PCR method. 【Background technique】 [0002] The function of eukaryotic translation initiation factor 1A (euka ryo tic translation initiation factor, eIF-1A) is mainly involved in the first step of the initiation of protein synthesis, that is, the composition of eIF2:GTP:tRNA with the participation of EIF1A, etc. A ternary complex that binds to the 40S ribosomal subunit to initiate translation. EIF1A plays an important role in the initiation of peptide chain synthesis and is one of the important regulators of protein synthesis. However, there is no research on the distribution of EIF1A gene in mango in different tissues. [0003] Fluorescent quantitative PCR (Real-time quantitative PCR, also known as quantitative PCR, referred to as QPCR) technology refers to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686C12Q1/6895C12Q1/6806C12N15/11C12N15/10
CPCC12N15/1003C12Q1/6806C12Q1/686C12Q1/6895C12Q2600/158C12Q2563/107C12Q2545/114
Inventor 李丽淑何新华罗聪
Owner GUANGXI UNIV
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