Characterization of reference genes for quantitative real-time PCR analysis in various tissues of Salvia miltiorrhiza.
Five reference genes, 18S, EF1alpha, alpha-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 EF1alpha 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.
Yang
Yanfang
Y
Institute of Medicinal Plant Development, Chinese Academy of Medicinal Sciences and Peking Union Medical College, Beijing, China.
Hou
Shuang
S
Cui
Guanghong
G
Chen
Shilin
S
Wei
Jianhe
J
Huang
Luqi
L
D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics*; D009928:Organ Specificity / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction; D027561:Salvia miltiorrhiza / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012984:Software; D014158:Transcription, Genetic
D010940:Plant Proteins
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