Identification of a novel reference gene for apple transcriptional profiling under postharvest conditions

PloS One
Tatiane Timm StorchCésar Luis Girardi

Abstract

Reverse Transcription quantitative PCR (RT-qPCR) is one of the most important techniques for gene expression profiling due to its high sensibility and reproducibility. However, the reliability of the results is highly dependent on data normalization, performed by comparisons between the expression profiles of the genes of interest against those of constitutively expressed, reference genes. Although the technique is widely used in fruit postharvest experiments, the transcription stability of reference genes has not been thoroughly investigated under these experimental conditions. Thus, we have determined the transcriptional profile, under these conditions, of three genes commonly used as reference--ACTIN (MdACT), PROTEIN DISULPHIDE ISOMERASE (MdPDI) and UBIQUITIN-CONJUGATING ENZYME E2 (MdUBC)--along with two novel candidates--HISTONE 1 (MdH1) and NUCLEOSSOME ASSEMBLY 1 PROTEIN (MdNAP1). The expression profile of the genes was investigated throughout five experiments, with three of them encompassing the postharvest period and the other two, consisting of developmental and spatial phases. The transcriptional stability was comparatively investigated using four distinct software packages: BestKeeper, NormFinder, geNorm and DataAssis...Continue Reading

References

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Citations

Mar 2, 2018·PloS One·Fabiane Igansi de Castro Dos SantosAntonio Costa de Oliveira

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Methods Mentioned

BETA
nucleic acid extraction
transcription profiling
PCR

Software Mentioned

Primer3Plus
BestKeeper
Normfinder
StepOne
DataAssist
geNorm

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