Internal reference gene for melon fruit genet PCR expression analysis, and stability verification method thereof

Abstract

The present invention discloses an internal reference gene for melon fruit genet PCR expression analysis, and a stability verification method for the internal reference gene. According to the present invention, the expression level and the stability of the gene are compared at the whole genome level by using melon transcriptome data so as to select eight stably-expressing genes, two traditional internal reference genes commonly used in melon are selected as controls, the expression stability of the 10 genes in two melon variety fruits at different development stages are compared, and it is determined that the expression stability of the MELO3C007745T1 gene is high highest through verification with genes such as Cmain2 and CmSPS1m, wherein the MELO3C007745T1 gene is the optimal internal reference gene for the real-time fluorescence quantitative PCR analysis in the melon fruit gene expression.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to an internal reference gene for PCR expression analysis of muskmelon fruit genes and a method for verifying the stability of the internal reference gene. Background technique [0002] Gene expression analysis is an important means to study gene function and transcriptional regulation. Real-time fluorescent quantitative PCR (qRT-PCR) is widely used in the quantitative analysis of gene expression because of its high sensitivity, high specificity, high accuracy and low cost. In qRT-PCR analysis, in order to obtain the real difference in the expression level of the target gene, it is necessary to eliminate the differences in the RNA yield, quality and reverse transcription efficiency of different samples. To eliminate the difference between samples, the internal reference gene is usually used for correction and standardization, that is, by measuring the expression o...

AI Technical Summary

Problems solved by technology

At present, most of the traditional internal reference genes are used in the gene expression analysis of melon. These traditional internal reference genes have not been systematically verified and have poor stability, resulting in poor accuracy of gene expression analysis in melon.

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