Identification and evaluation of reference genes for qRT-PCR studies in Lentinula edodes

PloS One
Quanju XiangYunfu Gu

Abstract

Lentinula edodes (shiitake mushroom) is a common edible mushroom with a number of potential therapeutic and nutritional applications. It contains various medically important molecules, such as polysaccharides, terpenoids, sterols, and lipids, were contained in this mushroom. Quantitative real-time polymerase chain reaction (qRT-PCR) is a powerful tool to analyze the mechanisms underlying the biosynthetic pathways of these substances. qRT-PCR is used for accurate analyses of transcript levels owing to its rapidity, sensitivity, and reliability. However, its accuracy and reliability for the quantification of transcripts rely on the expression stability of the reference genes used for data normalization. To ensure the reliability of gene expression analyses using qRT-PCR in L. edodes molecular biology research, it is necessary to systematically evaluate reference genes. In the current study, ten potential reference genes were selected from L. edodes genomic data and their expression levels were measured by qRT-PCR using various samples. The expression stability of each candidate gene was analyzed by three commonly used software packages: geNorm, NormFinder, and BestKeeper. Base on the results, Rpl4 was the most stable reference ge...Continue Reading

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Citations

Sep 15, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Qianqian ZhangYinbing Bian
Jan 4, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Shuangshuang GaoWen Huang
Jun 30, 2019·Journal of Microbiological Methods·Ding-Hong JiaWei-Hong Peng
Feb 16, 2021·The Journal of Veterinary Medical Science·Takuma AriizumiKazuhiko Ohashi
Oct 26, 2021·Journal of Agricultural and Food Chemistry·Yihong WangMingwen Zhao

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Methods Mentioned

BETA
electrophoresis
PCR

Software Mentioned

NormFinder
BestKeeper
Excel
- Blast
Primer
GeNorm
based
NorFinder

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