GSM8015651: JC2307 (DccrM Ptac-ccrM) ATGN+IPTG replicate C; Agrobacterium tumefaciens; RNA-Seq
Cells were pelleted from 1-2mL of bacterial cultures and immediately frozen in liquid nitrogen prior to conservation at -80°C. RNA were extracted using an RNeasy Mini-Kit from Qiagen and following the manufacturer's protocol and including a DNase I (RNase-free DNases set from Qiagen) treatment. Samples were additionally treated with a TURBO DNA-free kit from Invitrogen following the manufacturer's protocol. RNA-seq libraries were prepared from 800 ng of total RNA with the TruSeq Stranded mRNA Prep reagents (Illumina) using a unique dual indexing strategy, and following the official protocol automated on the Sciclone liquid handling robot (PerkinElmer). The polyA selection step was replaced by an rRNA depletion step with the QIAseq FastSelect - 5S/16S/23S bacterial rRNA removal kit (Qiagen).