Transcriptome Sequencing and De Novo Analysis for Youngia japonica Using the Illumina Platform
The mRNA-seq library was constructed following the manufacturer’s instructions of mRNA-Seq Sample Preparation Kit (Cat# RS-930-1001, Illumina Inc, San Diego, CA). The poly-(A) mRNA was isolated from the total RNA samples with poly-T oligo-attached magnetic beads.PCR was performed to selectively enrich and amplify the cDNA fragments using Phusion Master Mix (NEB) with two primers.The amplified products were purified according to the instructions of QIAquick PCR Purification Kit (QIAGEN) and eluted in 30 µL of QIAGEN EB buffer. Libraries were prepared from a 150-200 bp size-selected fraction following adapter ligation and agarose gel separation.