Evaluation of reference genes for quantitative polymerase chain reaction across life cycle stages and tissue types of Tribolium castaneum.
The genome of the genetic model for coleopteran insects, Tribolium castaneum, is now available for downstream applications. To facilitate gene expression studies in T. castaneum, genes were evaluated for suitability as normalizers in comparisons across tissues and/or developmental stages. In less diverse samples, such as comparisons within developmental stages or tissue only, normalizers for mRNA were more stable and consistent. Overall, the genes for ribosomal proteins rps6, rpl13a, rps3, and rps18 were the most stable normalizers for broad scale gene expression analysis in T. castaneum. However, their stability ranking was dependent upon the instrument as well as the analysis program. These data emphasize the need to optimize normalizers used in all real-time polymerase chain reaction experiments specifically for the experimental conditions and thermocycler and to carefully evaluate data generated by computational algorithms.
Toutges
Michelle J
MJ
Center for Grain and Animal Health Research, Agricultural Research Service, U.S. Department of Agriculture, 1515 College Avenue, Manhattan, Kansas 66502, USA. michelle.toutges@ars.usda.gov
Hartzer
Kris
K
Lord
Jeffrey
J
Oppert
Brenda
B
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D017344:Genes, Insect; D008018:Life Cycle Stages; D016133:Polymerase Chain Reaction / Q000379:methods*; D014231:Tribolium / Q000235:genetics* / Q000502:physiology
D017931:DNA Primers