RNA quality assessment: a view from plant qPCR studies

被引:43
作者
Die, Jose V. [1 ]
Roman, Belen [2 ]
机构
[1] Inst Agr Sostenible, Cordoba 14080, Spain
[2] IFAPA Alameda del Obispo, Biotecnol & Mejora, Cordoba 14080, Spain
关键词
MIQE; qPCR; 3 ':5 ' ratio; RNA integrity; REAL-TIME PCR; GENE-EXPRESSION ANALYSIS; LASER CAPTURE MICRODISSECTION; RT-PCR; MESSENGER-RNA; ARBUSCULAR MYCORRHIZAL; SYSTEMATIC VALIDATION; MINIMUM INFORMATION; DEGRADATION; IMPACT;
D O I
10.1093/jxb/ers276
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
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 ogold 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.
引用
收藏
页码:6069 / 6077
页数:9
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