title: RNA quality assessment: a view from plant qPCR studies.
abstract: Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) is probably the most common molecular technique used in transcriptome analyses today. The simplicity of the technology and associated protocols that generate results without the need to understand the underlying principles has made RT-qPCR the method of choice for RNA quantification. Rather than the 'gold standard technology' often used to describe it, the performance of RT-qPCR suffers from considerable pitfalls during general workflow. The inconsistency of conventional methods for the evaluation of RNA quality and its influence on qPCR performance as well as stability of reference genes is summarized and discussed here.
authors.affiliation: Mejora Genética Vegetal, Instituto de Agricultura Sostenible, CSIC, 14080 Córdoba, Spain. Jose.Die@ars.usda.gov
authors.forename: Jose V
authors.initials: JV
authors.lastname: Die
authors.forename: Belén
authors.initials: B
authors.lastname: Román
mesh_terms: D020869:Gene Expression Profiling / Q000592:standards; D010944:Plants / Q000235:genetics*; D018749:RNA, Plant / Q000032:analysis* / Q000378:metabolism / Q000592:standards; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards
chemical_list: D018749:RNA, Plant