GSM9678479: delta_HNS_MicroC_biol_rep2; Escherichia coli; OTHER
Cells were resuspended in 250 µL TE (10 mM Tris pH 8.0, 1 mM EDTA) supplemented with 1× protease inhibitors (Bimake) followed by adding lysozyme (Sigma L6876) to 4 µg/mL and incubating for 30 min at 37 °C with shaking at 1000 rpm. The sample was pelleted for 5 min at 16100 g and room temperature and resuspended in an appropriate volume of MNase buffer (10 mM Tris pH 7.5, 1 mM CaCl2) followed by adding MNase (ThermoFisher EN0181, used as 5 U/µL stock solution) to 10 U/mL and incubating for 25 min at 37 °C with shaking at 1000 rpm. The volume of MNase buffer ranged from 475 to 1160 µL for different samples; it was determined individually for each sample to obtain the fragment distribution from ~ 75 to 1000 bp. DNA digestion was terminated by adding EGTA to 5 mM followed by centrifugation for 5 min at 16100 g and room temperature. The libraries were prepared following a modified Micro-C protocol