title: RNA-seq of Brassica oleracea: leaf tissue 96h after infestation with (parasitised) aphids
design: Seven leaf samples from plants with the same treatment were pooled and RNA was extracted using the bioline kit. RNA quality was measured using a Bio-analyser (agilent). And RNA samples were shipped to BGI in Hongkong. Here, oligo dT beads were used to enrich mRNA containing a poly A tail.The mRNA molecules were fragmented into small pieces. First strand cDNA synthesis was performed using random primers on the fragmented mRNA.Subsequently, the second strand cDNA was synthesized. End-repair and 3' adenylation were carried out on the synthesized cDNA. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments. PCR amplification and purification were performed. The library's quality was assessed through library quality control measures. Library circularization was conducted.The library was then amplified to generate DNA nanoballs (DNBs). Sequencing was performed on the DNBs using the DNBSEQ Technology platform.