Qiagen RNeasy mini-prep kit (Qiagen, USA) was used for the extraction of RNA from about 3 ml of E. coli culture and cDNA synthesized using the first strand cDNA synthesis kit from Invitrogen (Invitrogen Bioservices India Pvt. Ltd., Bangalore) as described previously [12] (link). The cDNA synthesized for all the three replicates of LSP and SP E. coli cultures grown freshly in both LB broth and LB+glycerol broth was fragmented with DNAse 1 (Promega Corporation, Madison, USA) and then labelled with biotin at the 3′ end using the labelling reagent from Affymetrix (CA, USA) and Terminal transferase enzyme (Promega Corporation, Madison, USA).
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