GSM4816541: E. coli MG1655 Culture B Replicate 1; Escherichia coli str. K-12 substr. MG1655; RNA-Seq
10 mL of cell-culture was vacuum filtered using a Supor ® 800 Grid filter (Pall, NY, USA) with 0.8 µM pore size. The filter containing the cells was put in a 15 mL falcon tube containing 700 µL PGTX solution and immediately frozen in liquid nitrogen. The samples were stored at -80°C until further processing. RNA quality was determined using Bioanalyzer (Agilent, CA, USA). Ribo-Zero rRNA Removal Kit for Bacteria (Illumina, CA, USA) was used to deplete rRNA. First strand cDNA synthesis and second strand synthesis was carried out using NEBNext RNA First Stand and Second Strand Synthesis Module (New England Biolabs, MA, USA). cDNA was then purified and concentrated using MiniElute Columns (Qiagen, Netherlands) and the Encore Rapid DR Multiplex system (Tecan, Switzerland) was used for library preparation. Libraries were amplified for 12 cycles using MyTaq (Bioline, UK) and standard Illumina primers. Size selection was done using a preparative Agarose Gel selecting fragments between 300 and 500 bp and quality control of libraries was performed with Bioanalyzer and Qubit.
GEO Accession
GSM4816541