Whole transcriptome sequencing of the rhabodvirus-negative Sf9 cell clone 13F12
Cellular RNA analyzed was processed to generate libraries for mRNA sequencing following the Illumina® TruSeq Stranded mRNA Sample Preparation Guide .). In this method, polyA containing mRNA was purified from 1 µg total RNA using poly-T oligo attached magnetic beads, fragmented and reverse-transcribed into cDNAs.Double strand cDNAs were adenylated at the 3' ends and ligated to indexed sequencing adaptors, followed with briefly amplification for 15 cycles.  The quality of all cDNA libraries was assessed by the Agilent 2100 Bioanalyzer using the DNA 1000 Kit (Agilent). One femtomole of the sequencing libraries (median size ~260nt) were denatured and loaded onto a flow cell for cluster generation using the Illumina cBot. Each sample was loaded onto each lane of a HiSeq Rapid Run flow cell. Paired-end sequencing was carried out on HiSeq 2500 sequencer (Illumina, San Diego, CA, USA) for 100 x 2 cycles. For each sample, we obtained ~100 million 100-bp reads that passed preset filtering parameters.