Illumina RNA-Seq Transcriptome Sequencing

Total RNA was extracted from 9 tissues (5 FD stages and 4 plant parts) and cDNA libraries were prepared. RNA quality and quantity check was made through Nano-Drop spectrophotometer and cDNA libraries were prepared according to Illumina protocol [48 (link)]. The mRNA fragments of about 200 bp were obtained from total RNA, and first and second strand cDNAs were prepared and adapter ligation and low-cycle enrichment were performed according to TruSeq^®RNA HT Sample Prep Kit from Illumina (San Diego, CA, USA). The purified library products were diluted to 10 pM, and sequencing was performed. Finally, de novo transcriptome was done with Trinity program using default parameters [49 (link)].