The Verification of Differentially Expressed Genes Related to Feather Colour in Quail Embryos by RT-qPCR Based on RNA-seq

Abstract

Many differentially expressed genes associated with feather colour in quail embryos can be screened using RNA-seq technology. The aim of this study was to verify the accuracy of RNA-seq using RT-qPCR. The transcriptome of embryonic skin tissues of quails were sequenced by Illumina HiSeq TM2000 sequencing platform. As a result, 2512 differentially expressed genes were found. Among them, 950 differentially expressed genes were up-regulated in the skin tissues of white-feather quail embryos, and 1562 differentially expressed genes were down-regulated. The candidate genes were analyzed by RT-qPCR method to verify the accuracy and reliability of the RNA-seq. The results showed that the relative expression levels of ASIP and DDC in white-feather quails were significantly increased, while HOXD1, CTSD, KRT2, MBD2 and MT1 were significantly decreased, which was consistent with the results of RNA-seq. The validation results of the candidate genes can also provide some ideas to explain the difference in feather colour between maroon-feather quails and white-feather quails.