Transcriptome sequencing of Streptomyces sp. AgN23

AgN23 was cultivated in liquid Bennett medium at 28 C for 48h. Total RNA was isolated using the RNeasy Plant Mini Kit (Qiagen) according to manufacturer instructions. Three replicates were prepared for the libraries construction. rRNA depletion was performed using Ribozero rRNA Removal Kit (Illumina). RNA sequencing library preparation used NEBNext Ultra RNA Library Prep Kit for Illumina by following the manufacturer recommendations (NEB). RNAs were fragmented to generate double stranded cDNA, subsequently ligated to universal adapter, followed by index addition and library enrichment with limited cycle PCR. Sequencing libraries were validated using the Agilent Tapestation 4200 (Agilent Technologies), and quantified by Qubit 2.0 Fluorometer (Invitrogen) as well as quantitative PCR (Applied Biosystems). RNA-Seq experiments have been performed on an Illumina HiSeq 4000 using a paired-end read length of 2x150 bp with the Illumina HiSeq 4000 sequencing kits.