| Literature DB >> 32485234 |
Alexander P Schwarz1, Daria A Malygina2, Anna A Kovalenko2, Alexander N Trofimov3, Aleksey V Zaitsev2.
Abstract
RT-qPCR requires an adequate choice of stably expressed reference genes for accurate normalization of mRNA expression. However, testing a panel of reference genes is often time-consuming and expensive. In this work, we aimed to develop a set of multiplex real-time PCR assays for RT-qPCR analysis of commonly used housekeeping genes in laboratory rats. Using Hydrolysis probe (TaqMan®) technology, we have designed and optimized three triplex qPCR assays (Actb + Gapdh + B2m; Rpl13a + Sdha + Ppia; Hprt1+Pgk1+Ywhaz) demonstrating optimal PCR amplification efficiencies (from 94.7 to 100.5%) and repeatability. Novel assays allow expression analysis of 9 reference genes in 3 reactions making possible a more time-efficient choice of reference genes in RT-qPCR experiments in Wistar rats in comparison with widespread singleplex assays.Entities:
Keywords: Gene expression analysis; Multiplex real-time PCR assay; RT-qPCR; Rat; Reference gene stability
Mesh：
Year: 2020 PMID： 32485234 DOI： 10.1016/j.mcp.2020.101611
Source DB: PubMed Journal: Mol Cell Probes ISSN： 0890-8508 Impact factor: 2.365