Exogenous spike-in mouse RNAs for accurate differential gene expression analysis in barley using RT-qPCR.

ABSTRACT: Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) followed by the 2^-ΔΔCt method is the most common way to measure transcript levels for relative gene expression assays. The quality of an RT-qPCR assay is dependent upon the identification and validation of reference genes to normalize gene expression data. The so-called housekeeping genes are commonly used as internal reference genes because they are assumed to be ubiquitously expressed at stable levels. Commonly, researchers do not validate their reference genes but rely on historical reference genes or previously validated genes from an unrelated experiment. Using previously validated reference genes to assess gene expression changes occurring during malting resulted in extensive variability. Therefore, a n

SUBMITTER: Vinje MA 

PROVIDER: S-EPMC10728042 | biostudies-literature | 2023

REPOSITORIES: biostudies-literature