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Characterization of reference genes for quantitative real-time PCR analysis in various tissues of Salvia miltiorrhiza

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Abstract

Five reference genes, 18S, EF1α, α-Tubulin, Ubiquitin and Actin, from Salvia miltiorrhiza were analyzed as internal controls for gene expression profiling assay using quantitative real-time polymerase chain reaction (qRT-PCR). The five candidate genes were measured for their transcriptional level in seven tissues, including roots, stems, leaves, sepals, petals, stamens and pistils. Then they were ranked by the GeNorm tool. The results showed that Actin and Ubiquitin were the most stable whereas EF1α and 18S did not favor normalization of qRT-PCR results in these tissues. Expression levels of the SmDXR gene were studied in parallel, with Actin and Ubiquitin both or each as reference in the seven tissues, and varying relative quantifications of the SmDXR gene in seven tissues. This study indicated that selection of the most stable genes plays an important role in gene expression profiling assays.

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Abbreviations

DMAPP:

Dimethylallyl diphosphate

DXR:

1-Deoxy-d-xylulose 5-phosphate reductoisomerase

IPP:

Isopentenyl diphosphate

MEP:

2-c-Methyl-d-erythritol 4-phosphate

MVA:

Mevalonate acid

PCR:

Polymerase chain reaction

qRT-PCR:

Quantitative real-time PCR

RT:

Reverse transcription

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Acknowledgments

This work was supported by The Special Funds in Basic Scientific Research for Non-Profit Research Institutes financed by the Ministry of Finance People’s Republic of China (No. YZ-08-19).

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Authors and Affiliations

  1. Institute of Medicinal Plant Development, Chinese Academy of Medicinal Sciences and Peking Union Medical College, NO.151, Malianwa North Road, 100193, Hai Dian District, Beijing, China

    Yanfang Yang, Shuang Hou, Shilin Chen & Jianhe Wei

  2. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, 100700, Beijing, China

    Guanghong Cui & Luqi Huang

  3. College of Horticultural and Landscape Architecture, Southwest University, 400715, Chongqing, China

    Shuang Hou

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  1. Yanfang Yang
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  2. Shuang Hou
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Correspondence to Jianhe Wei or Luqi Huang.

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Yang, Y., Hou, S., Cui, G. et al. Characterization of reference genes for quantitative real-time PCR analysis in various tissues of Salvia miltiorrhiza . Mol Biol Rep 37, 507–513 (2010). https://doi.org/10.1007/s11033-009-9703-3

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