GSM5900568: ItaT; Escherichia coli str. K-12 substr. MG1655; OTHER
The cells were harvested and lyzed as described in Becker et al. 2013. The cellular lysates were clarified by centrifugation at 20,000 x g at 4°C for 10 min. The lysates were diluted with the lysis buffer  to adjust the OD260 to 0.1 in the final volume of 220 μl and treated with ~450 U of MNase (Roche) at 25°C for 1 hr. The reaction was quenched by the addition of EGTA to the final concertation of 5 mM. The monosomal material was purified by sucrose gradient centrifugation. RNA was purified as described in Becker et al. 2013. Libraries were constructed as described in Becker et al. 2013