Department of Life Sciences and Systems Biology, University of Torino, Torino, Italy.
Department of Soil Microbiology and Symbiotic Systems, Estación Experimental del Zaidín (EEZ-CSIC), Granada, Spain.
Methods Mol Biol. 2020;2146:157-170. doi: 10.1007/978-1-0716-0603-2_12.
Gene expression analysis is a broadly used and powerful technique in many fields of biological research. The expression pattern of specific marker genes provides an insight into complex regulatory networks and leads to the identification of relevant genes associated to specific biological processes, such as arbuscular mycorrhizal symbiosis. Among the existing gene expression analysis toolbox, reverse transcriptase coupled to quantitative polymerase chain reaction (qRT-PCR) is considered the gold standard for accurate, sensitive, fast, and relatively inexpensive measurement. However, for a correct identification of differentially expressed genes, appropriate controls are required in order to minimize nonspecific variations associated with intrinsic technical variability. In this chapter, we recommend a number of tips to use qRT-PCR analysis in mycorrhizal roots and fungal mycelium.
基因表达分析是生物研究众多领域中广泛使用且功能强大的技术。特定标记基因的表达模式提供了对复杂调控网络的深入了解，并导致与特定生物过程相关的相关基因的鉴定，例如丛枝菌根共生。在现有的基因表达分析工具箱中，逆转录酶定量聚合酶链反应（qRT-PCR）被认为是准确、灵敏、快速且相对廉价的测量的金标准。然而，为了正确识别差异表达的基因，需要适当的对照，以最小化与内在技术变异性相关的非特异性变化。在本章中，我们推荐了一些在菌根根和真菌菌丝中使用 qRT-PCR 分析的技巧。
