Evaluation of Appropriate Reference Genes for Reverse Transcription-Quantitative PCR Studies in Different Tissues of a Desert Poplar via Comparision of Different Algorithms

International Journal of Molecular Sciences
Hou-Ling WangXinli Xia

Abstract

Despite the unshakable status of reverse transcription-quantitative PCR in gene expression analysis, it has certain disadvantages, including that the results are highly dependent on the reference genes selected for data normalization. Since inappropriate endogenous control genes will lead to inaccurate target gene expression profiles, the validation of suitable internal reference genes is essential. Given the increasing interest in functional genes and genomics of Populus euphratica, a desert poplar showing extraordinary adaptation to salt stress, we evaluated the expression stability of ten candidate reference genes in P. euphratica roots, stems, and leaves under salt stress conditions. We used five algorithms, namely, ΔCt, NormFinder, geNorm, GrayNorm, and a rank aggregation method (RankAggreg) to identify suitable normalizers. To support the suitability of the identified reference genes and to compare the relative merits of these different algorithms, we analyzed and compared the relative expression levels of nine P. euphratica functional genes in different tissues. Our results indicate that a combination of multiple reference genes recommended by GrayNorm algorithm (e.g., a combination of Actin, EF1α, GAPDH, RP, UBQ in root...Continue Reading

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Citations

May 20, 2017·Frontiers in Plant Science·Xue ChenLifang Wu
Aug 31, 2019·International Journal of Molecular Sciences·Valentin AmbroiseKjell Sergeant

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Methods Mentioned

BETA
environmental stress
PCR
electrophoresis
salt treatment
salt stress

Software Mentioned

RankAggreg
geNorm
NormFinder
NormFinder Basic Application ( VBA
GenEx
SPSS
R
qBasePlus
Excel
GrayNorm

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