Reference Gene Selection for Insect Expression Studies Using Quantitative Real-Time PCR: The Head of the Honeybee,Apis mellifera, After a Bacterial Challenge

• DOI: 10.1673/031.008.3301
• Publication Date: 2009-06-25T04:59:08Z
• Authors: Bieke Scharlaken; Dirk C. de Graaf; Karen Goossens; Marleen Brunain; Luc J. Peelman; Frans J. Jacobs
• Keywords: Reference Genes; Candidate gene

In this study an important and often neglected aspect of gene expression studies in insects, the validation appropriate reference genes with stable levels between sample groups, is addressed. Although paper selection for honeybee, Apis mellifera L. (Hymenoptera: Apidae) head was tested context bacterial challenge Escherichia coli, work can serve as a resource to help select screen insect any tissue under experimental manipulation. Since it recommended use multiple accurate normalization, we analyzed eleven candidate honeybee head, their potential analysis differential following challenge. Three software programs, BestKeeper, Normfinder geNorm, were used assess genes. GeNorm four Both geNorm identified GAPDH, RPS18, actin RPL13a most ones, only differing ranking order. BestKeeper RPS18 being least overall variation, but also GAPDH found be second third expressed gene. By combination three programs actin, suitable infection.