Illumina HiSeq 2000 sequencing; Transcriptome before and 10 min after valine induced isoleucine starvation in Escherichia coli K12 grown in minimal MOPS glucose medium at 37° C
Transcriptome before and 10 min after valine induced isoleucine starvation in Escherichia coli K12 grown in minimal MOPS glucose medium at 37° C
Cells harvested right before and 10 min after valine addition. 280 ml of cultures were harvested, centrifuged and washed in 0° C biotin-free medium 10 generations of growth on MOPS minimal medium supplemented with 0.2% glucose and 5 µg/ml biotin Isoleucine starvation introduced by addition of 400 µg/ml valine Following collection, resuspension in 2 ml of lysis buffer, followed by sonication. Phenol extraction of aliquote followed by storage at -80°C libraries constructed WITHOUT rRNA depletion by 1) fragmentation of RNA 2) first strand synthesis with random primers 3) second strand synthesis with random primers 4) end repair and A tailing 5) adapter ligation 6) size selection and quantification and 7) sequencing
Experimental Factor: valine
compound
Experimental Factor: 400
dose