GSM7158028: Ara_PtuAB_rep2; Escherichia coli; RNA-Seq
Total RNA was extracted using miRNA isolation kit (QIAGEN, catalog number 217004), according to the manufacturer's recommendation. Over 2g total RNA was resolved on Urea-PAGE and recovered within a size window of 60-100 nt for tRNA. Thereafter, purified tRNA was demethylated, alkaline-hydrolyzed by carbonate buffer, dephosphorylated by calf intestinal phosphatase (CIP), and re-phosphorylated by T4 polynucleotide kinase (T4 PNK). Fragments of 19-35nt for partially hydrolyzed and re-phosphorylated tRNA were converted to small RNA sequencing libraries using NEBNext Multiplex Small RNA Library Prep Set for Illumina kit. tRNA-seq