RT Journal Article
ID 60f3ee9239ed0c56
A1 Yang, Chi
A1 Ma, Lu 
A1 Xiao, Donglai 
A1 Ying, Zhenghe 
A1 Jiang, Xiaoling 
A1 Lin, Yanquan
T1 Identification and Evaluation of Reference Genes for qRT-PCR Normalization in <i>Sparassis latifolia</i> (Agaricomycetes)
JF International Journal of Medicinal Mushrooms
JO IJM
YR 2019
FD 2019-03-19
VO 21
IS 3
SP 301
OP 309
K1 normalization
K1 real-time quantitative polymerase chain reaction
K1 reference genes
K1 <i>Sparassis latifolia</i>
K1 light
K1 edible and medicinal mushrooms
AB Real-time quantitative polymerase chain reaction (qRT-PCR) has emerged as a powerful and popular tool for quantitating differences in transcriptional gene expression levels between samples. Validation of the stability of reference genes is a fundamental step before initiating qRT-PCR assays. <i>Sparassis latifolia</i> is an edible and medicinal fungus containing a remarkably high concentration of &beta;-glucan, which has many biological and pharmacologic activities. <i>S. latifolia</i> may be a model species for studying fungal photobiology because its fruiting body formation requires more light than other fungi. However, suitable reference genes for qRT-PCR have not yet been determined. In the present study, 10 candidate reference genes in <i>S. latifolia</i> were evaluated and validated under different developmental stages and light conditions. To evaluate the suitability of candidate reference genes, three popular software programs (geNorm, NormFinder, and BestKeeper), along with the delta Ct method, were used to analyze these genes; the final ranking was determined using RefFinder. According to our results, Actin and GAPDH were expressed at the most stable levels under different developmental stages and light conditions.

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