tailieunhanh - báo cáo khoa học: " Validation of reference genes for quantitative real-time PCR during leaf and flower development in Petunia hybrida"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Validation of reference genes for quantitative real-time PCR during leaf and flower development in Petunia hybrida | Mallona et al. BMC Plant Biology 2010 10 4 http 1471-2229 10 4 BMC Plant Biology RESEARCH ARTICLE Open Access Validation of reference genes for quantitative real-time PCR during leaf and flower development in Petunia hybrida Izaskun Mallona 1 Sandra Lischewski2 Julia Weiss1 Bettina Hause2 Marcos Egea-Cortines1 Abstract Background Identification of genes with invariant levels of gene expression is a prerequisite for validating transcriptomic changes accompanying development. Ideally expression of these genes should be independent of the morphogenetic process or environmental condition tested as well as the methods used for RNA purification and analysis. Results In an effort to identify endogenous genes meeting these criteria nine reference genes RG were tested in two Petunia lines Mitchell and V30 . Growth conditions differed in Mitchell and V30 and different methods were used for RNA isolation and analysis. Four different software tools were employed to analyze the data. We merged the four outputs by means of a non-weighted unsupervised rank aggregation method. The genes identified as optimal for transcriptomic analysis of Mitchell and V30 were EF1a in Mitchell and CYP in V30 whereas the least suitable gene was GAPDH in both lines. Conclusions The least adequate gene turned out to be GAPDH indicating that it should be rejected as reference gene in Petunia. The absence of correspondence of the best-suited genes suggests that assessing reference gene stability is needed when performing normalization of data from transcriptomic analysis of flower and leaf development. Background The general aims of transcriptomic analysis are identification of genes differentially expressed and measurement of the relative levels of their transcripts. Transcriptomic analysis like that relying on microarray techniques reveals an underlying expression dynamic that changes between tissues and over time 1 . Results must then be validated by other means in order to .