GSM9308078: mock inoculation (control), 2 days post-inoculation, rep2; Hevea brasiliensis; RNA-Seq
Leaf samples for each treatment were pooled, snap-frozen in liquid nitrogen, ground into a fine powder, and stored at -80 °C. Total RNA was isolated using the CTAB buffer method. RNA concentration, purity, and integrity were assesed using a spectrophotometer (Eppendorf BioSpectrometer Basic), electrophoresis in 1% agarose gel. At least 400 ng of total RNA was used to RNA-Seq. The oligo dT beads was used to enrich mRNA with poly A tail. The fragments mRNA was synthesized into first strand cDNA using random primers and the second strand cDNA was synthesized with dUTP instead of dTTP. The synthesized cDNA was subjected to end-repair and 3' adenylated. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments.