Morus alba Transcriptome NS7
Total RNA was isolated from preserved mulberry leaves after 7 days post treatment. Total RNA was used to prepare the RNA seq library using the TruSeq RNA Sample Prep Kits (Illumina). The polyA containing mRNA molecules were purified using polyT oligo attached magnetic beads. Following purification, the mRNA was fragmented into small pieces using divalent cations under elevated temperature. The cleaved RNA fragments were used to synthesize first strand cDNA using reverse transcriptase and random primers followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. These cDNA fragments were gone through an end repair process, the addition of a single A base, and then ligation of the adapters. The products were purified and enriched with PCR to create the final cDNA library. Bioanalyzer plots were used at every step to assess mRNA quality, enrichment success, fragmentation sizes and final library sizes. The size distribution of the sequencing library was determined by gel electrophoresis. Qubit was used for measuring the quantity of the library before sequencing. After the libraries were constructed, paired end run was performed on Illuminas HiSeq 4000 platform to obtain 2 x100bp reads.