title: Illumina HiSeq 2500 sequencing; Differential expression of E. coli(pBIC-1a) under imipenem exposure
design: Differential expression of E. coli(pBIC-1a) under imipenem exposure
library_construction_protocol: An overnight culture of E. coli TOP10-pBIC1a was diluted 100-fold in fresh BHI broth and allowed to grow to an OD600 of 0.5. The culture was then split and imipenem (Imipenem-Cilastatin, Mylan, USA) was added to one of the cultures at a final concentration of 5 µg/ml. Ten minutes later, 2 ml of each culture were taken and mixed with RNA protect (Qiagen, Courtaboeuf, France) according to the manufacturer's recommendations. Cells were immediately pelleted by a 10 min centrifugation at 5000 g.  RNA was extracted using GeneJet RNA extraction kit (ThermoScientific). DNAse treatment was performed with the dsDNAse kit (ThermoScientific) cDNA libraries were prepared using the NEBNext® Ultra Directional RNA Library Prep Kit for Illumina protocol (New England Biolabs, MA, USA). The quality of the cDNA was validated using the Agilent 2100 Bioanalyzer DNA1000 kit (Agilent Technologies) and quantity was determined with the Qubit dsDNA BR Assay (ThermoFisher Scientific).
attributes.tag: Experimental Factor: compound
attributes.value: imipenem
attributes.tag: Experimental Factor: dose
attributes.value: 5