alias: GSM2122747
title: GSM2122747: Erythromycin_replicate_2; Escherichia coli; RNA-Seq
library_construction_protocol: Briefly, hot phenol-chloroform extraction was done by mixing the cell culture in ½ volume of 99°C lysis solution (2% SDS, 16 mM EDTA, 200 mM NaCl made in RNAse-free H2O) for 10 min. The suspension was extracted twice with 1 volume of 65°C acid phenol/chloroform (pH 4.5), once in chloroform/isoamyl alcohol, and precipitated with isopropanol. Pellets were washed with 1 mL of 70% ethanol, dried and purified using QIAGEN RNeasy kit. Final RNA was eluted with 100 µL of nuclease-free water. Following purification, DNase treatment with TURBO DNase kit was performed according to manufacturer’s manual, except that twice amount of recommended DNase inactivation reagent was used. The integrity of DNA-free RNA was analyzed on either native agarose gel or on Agilent Bioanalyzer RNA 6000 Nano chip. Ion Xpress barcoded libraries were constructed using the Ribo-Zero Magnatic Kit (Gram-negative bacteria, Epicentre )and IonXpress RNA-seq v2 (Life Technologies) kits according to the manufacturer's directions.
identifiers.namespace: GEO
identifiers.id: GSM2122747