GSM2069220: Dodecanol Replicate 2; Acinetobacter venetianus RAG-1 = CIP 110063; RNA-Seq
The extracted RNA was subjected to Turbo DNase to get rid of any DNA contamination. The RNA obtained was analyzed using Agilent 2100 Bioanalyzer with the Agilent RNA 6000 Nano Kit. The RIN (RNA Integrity Number) values of all samples were above 9.6 indicating high quality RNA. The total RNA samples were prepared for Illumina Next-Generation Sequencing at the Functional Genomics Lab (FGL, QB3-Berkeley Core Research Facility at University of California, Berkeley). RiboZero kit (Illumina) was used to deplete ribosomal RNA. The treated RNA was re-checked on Bioanalyzer for integrity. The library preparation was performed on the Apollo 324TM with PrepXTM RNA-Seq Library Preparation Kit (WaferGen Biosystems, Fremont, CA) according to the manufacturer’s recommendation. The resulting samples were quantified using fluorimetry (Qubit, ThermoFischer) and PCR amplified for 18 cycles to incorporate indexes and flow cell-binding regions. Final libraries were quantified by Qubit, Bioanalyzer and qPCR, pooled at 14 picomolar concentration and sequenced on an Illumina HiSeq2000 using 50 base single read version 3 chemistry.