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Selection and Validation of Reference Genes for Quantitative Real-time PCR in Gentiana macrophylla

Overview of attention for article published in Frontiers in Plant Science, June 2016
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Title
Selection and Validation of Reference Genes for Quantitative Real-time PCR in Gentiana macrophylla
Published in
Frontiers in Plant Science, June 2016
DOI 10.3389/fpls.2016.00945
Pubmed ID
Authors

He, Yihan, Yan, Hailing, Hua, Wenping, Huang, Yaya, Wang, Zhezhi

Abstract

Real time quantitative PCR (RT-qPCR or qPCR) has been extensively applied for analyzing gene expression because of its accuracy, sensitivity, and high throughput. However, the unsuitable choice of reference gene(s) can lead to a misinterpretation of results. We evaluated the stability of 10 candidates - five traditional housekeeping genes (UBC21, GAPC2, EF-1α4, UBQ10, and UBC10) and five novel genes (SAND1, FBOX, PTB1, ARP, and Expressed1) - using the transcriptome data of Gentiana macrophylla. Common statistical algorithms ΔC t, GeNorm, NormFinder, and BestKeeper were run with samples collected from plants under various experimental conditions. For normalizing expression levels from tissues at different developmental stages, GAPC2 and UBC21 had the highest rankings. Both SAND1 and GAPC2 proved to be the optimal reference genes for roots from plants exposed to abiotic stresses while EF-1α4 and SAND1 were optimal when examining expression data from the leaves of stressed plants. Based on a comprehensive ranking of stability under different experimental conditions, we recommend that SAND1 and EF-1α4 are the most suitable overall. In this study, to find a suitable reference gene and its real-time PCR assay for G. macrophylla DNA content quantification, we evaluated three target genes including WRKY30, G10H, and SLS, through qualitative and absolute quantitative PCR with leaves under elicitors stressed experimental conditions. Arbitrary use of reference genes without previous evaluation can lead to a misinterpretation of the data. Our results will benefit future research on the expression of genes related to secoiridoid biosynthesis in this species under different experimental conditions.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 25 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Serbia 1 4%
Unknown 24 96%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 9 36%
Student > Bachelor 3 12%
Student > Doctoral Student 2 8%
Professor 2 8%
Student > Master 1 4%
Other 2 8%
Unknown 6 24%
Readers by discipline Count As %
Agricultural and Biological Sciences 12 48%
Biochemistry, Genetics and Molecular Biology 3 12%
Environmental Science 1 4%
Immunology and Microbiology 1 4%
Unknown 8 32%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 25 July 2016.
All research outputs
#17,810,002
of 22,880,230 outputs
Outputs from Frontiers in Plant Science
#12,078
of 20,270 outputs
Outputs of similar age
#252,793
of 352,012 outputs
Outputs of similar age from Frontiers in Plant Science
#267
of 526 outputs
Altmetric has tracked 22,880,230 research outputs across all sources so far. This one is in the 19th percentile – i.e., 19% of other outputs scored the same or lower than it.
So far Altmetric has tracked 20,270 research outputs from this source. They receive a mean Attention Score of 4.0. This one is in the 31st percentile – i.e., 31% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 352,012 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 24th percentile – i.e., 24% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 526 others from the same source and published within six weeks on either side of this one. This one is in the 40th percentile – i.e., 40% of its contemporaries scored the same or lower than it.