selection and validation of reference genes for quantitative real-time pcr in buckwheat (fagopyrum esculentum) based on transcriptome sequence data选择和验证的参考基因定量实时pcr在荞麦(fagopyrum esculentum)基于转录组序列数据.pdfVIP

Selection and Validation of Reference Genes for Quantitative Real-Time PCR in Buckwheat (Fagopyrum esculentum) Based on Transcriptome Sequence Data 1 2,3,4 1,3,4 Natalia V. Demidenko , Maria D. Logacheva , Aleksey A. Penin * 1 Department of Genetics, Biological Faculty, M.V. Lomonosov Moscow State University, Moscow, Russia, 2 Department of Evolutionary Biochemistry, A.N. Belozersky Institute of Physico-Chemical Biology, M.V. Lomonosov Moscow State University, Moscow, Russia, 3 Evolutionary Genomics Laboratory, Faculty of Bioengineering and Bioinformatics, M.V. Lomonosov Moscow State University, Moscow, Russia, 4 A.A. Kharkevich Institute for Information Transmission Problems, Russian Academy of Science, Moscow, Russia Abstract Quantitative reverse transcription PCR (qRT-PCR) is one of the most precise and widely used methods of gene expression analysis. A necessary prerequisite of exact and reliable data is the accurate choice of reference genes. We studied the expression stability of potential reference genes in common buckwheat (Fagopyrum esculentum) in order to find the optimal reference for gene expression analysis in this economically important crop. Recently sequenced buckwheat floral transcriptome was used as source of sequence information. Expression stability of eight candidate reference genes was assessed in different plant structures (leaves and inflorescences at two stages of development and fruits). These genes are the orthologs of Arabidopsis genes identified as stable in a genome-wide survey gene of expression stability and a traditionally used