Normalization of qPCR in platelets – YWHAE a potential genericreference gene
作者： Karin Mossberg , Per-Arne Svensson , Olof Gidlöf , David Erlinge , Sverker Jern
DOI: 10.1080/09537104.2016.1180349
关键词:
摘要: The mRNA of human platelets has been extensively studied and it is generally appreciated that contain transcripts derived from the megakaryocytes, they have ability to translate into proteins. Additionally, microRNA (miRNA) shown potentially regulate translation certain When quantifying gene expression by quantitative real-time polymerase chain reaction (qPCR), a valid normalization method required use reference genes common robust approach. It recommended perform proper validation potential for each individual experimental setup. Previous studies mainly performed using commonly used nucleated cells, our knowledge there are no global evaluations stability in platelets. Finding stable transcript would be valuable inter-study comparisons, aim this study was identify one or more suitable as generic Platelets were incubated 24 h microarray platelet revealed levels YWHAE, B2M, ITM2B, H3F3A, PF4V1 remained similar between 0 h. Further these together with GAPDH, RN18S1, PPIA, frequently studies, qPCR after different vitro conditions. In addition, inter-individual analyzed diabetic patients compared healthy matched controls. Analysis software RefFinder PF4V1, B2M most donors. YWHAE subjects. Furthermore, influence miRNA on selected investigated knockdown Dicer1 megakaryocytic cell line MEG01. GAPDH unchanged over time MEG01 cells indicating not regulated hence stably expressed. conclusion, we suggest studies.
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