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Evaluation of suitable reference genes for normalization of microRNA expression by real-time reverse transcription PCR analysis during longan somatic embryogenesis

Lin, Yu Ling ; Lai, Zhong Xiong

Plant Physiology and Biochemistry, May 2013, Vol.66, pp.20-25 [Peer Reviewed Journal]

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  • Title:
    Evaluation of suitable reference genes for normalization of microRNA expression by real-time reverse transcription PCR analysis during longan somatic embryogenesis
  • Author: Lin, Yu Ling ; Lai, Zhong Xiong
  • Description: Accurate profiling of microRNAs (miRNAs) is an essential step for understanding both developmental and physiological functions of miRNAs. Real-time quantitative PCR (qPCR) is being widely used in miRNA expression studies, but choosing a suitable reference gene is a crucial factor for correct analysis of results. To date, there has been no systematic evaluation of qPCR reference genes for the study of miRNAs during somatic embryogenesis (SE) in the longan tree ( ). Here, the most stably expressed miRNAs in synchronized longan tree embryogenic cultures at different developmental stages were determined using the geNorm and NormFinder algorithms. Validation qPCR experiments were performed for 24 miRNAs together with a snRNA ( ), a rRNA ( ), and three housekeeping genes. It was found that small RNAs had better expression stability than protein-coding genes, and was identified as the most reliable reference gene, followed by , and . was recommended as a normalizer if only a single reference gene was to be used, while the combination of , and was preferred to normalize miRNA expression data during longan SE. ► Expression stability of longan RNAs was compared using geNorm and NormFinder. ► Small RNAs had better expression stability than protein-coding genes. ► was recommended if only a single reference gene was to be used. ► A combination of three small RNAs was preferred to normalize miRNA expression.
  • Is Part Of: Plant Physiology and Biochemistry, May 2013, Vol.66, pp.20-25
  • Identifier: ISSN: 0981-9428 ; E-ISSN: 1873-2690 ; DOI: 10.1016/j.plaphy.2013.02.002
  • Subjects: Dimocarpus Longan ; Somatic Embryogenesis ; Microrna ; Reference Genes ; Qpcr ; Normalization ; Botany ; Chemistry
  • Language: English
  • Source: ScienceDirect Journals (Elsevier)

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