title: RNA-Seq of BBTV-NonViruliferous Pentalonia nigronervosa:Whole body: reared on BBTV free banana for the first read
design: A. RNA isolation, library preparation and RNA-seqBoth Viruliferous (Vr) and Non Viruliferous (NVr) banana aphid samples were RNA extracted using RNeasy Plus kit (Qiagen, MD, USA) ccording to the manufacturers instructions. The quantity and quality of the total RNA were validated using NanoDrop for the purity of the RNA samples, and Agilent 2100 Bioanalyzer ( Agilent RNA 6000 Nano Kit) for the RNA integrity (RIN value), 28S/18S and the fragment length distribution. The samples were further sequenced using BGISEQ-500 platform following the workflow in brief as follow; 1)mRNA enrichment, 2) RNA fragment and reverse transcription: 3)End repair, 4) PCR amplification, 5)Denature and cyclization, 6)Sequencing on BGISEQ-  500 platform. B. RNA-seq data workflowA quality control step was first performed on the raw sequencing reads. Main workflow for filtering the raw read includes; 1) Removing the  reads with adaptors; 2) Removing the  reads in which unknown bases(N) are more than 5%; 3) Removing low quality reads (low quality was define as the read where the percentage of base quality is lesser than 15 is greater than 20% in a read). After filtering, the remaining reads are called "Clean Reads" and stored in FASTQ file. Those clean read then ready for assembly