|Application:||Gene expression microarray analysis|
|Number of samples:||12|
|Release date:||Aug 31 2005|
|Last update date:||Mar 16 2012|
|Dataset link||Effect of small RNA RyhB on global iron use in Escherichia coli|
Derivatives of MG1655 carrying either pBAD-ryhB or the control vector pNM12 were used in all experiments. The strains used contain the (delta)ara714 allele to prevent catabolism of arabinose and the (delta)ryhB::cat allele to restrict RyhB expression to the inducible pBAD-ryhB vector. Overnight bacterial cultures were incubated in LB media with ampicillin at a final concentration of 50 ug/mL at 37oC and diluted 1000-fold into 50 mL of fresh LB-ampicillin media at 37oC with agitation. To induce RyhB expression, cultures carrying the pBAD-ryhB construct were grown to an O.D.600 of 0.5 and arabinose was added to the culture at a final concentration of 0.1%. In some experiments, 50 uM FeSO4 was added to the new culture after dilution from overnight culture. Total RNA was extracted from cells at the indicated time using the hot phenol procedure.