title: GSM9167573: M. xanthus TRIC-seq, stationary; Myxococcus xanthus; OTHER
library_name: GSM9167573
library_construction_protocol: Cells were pelleted, and crosslinks were reversed by incubation with Proteinase K at 65°C for 3 hours. Total RNA was extracted using PCI (Phenol:Chloroform:Isoamyl alcohol for RNA, Carl Roth, #X985.3). The biotinylated chimeric RNAs were captured using MyOne Streptavidin C1 beads (Thermo Fisher, #65002) The construction of TRIC-seq libraries includes a multi-step in situ protocol involving cell fixation, permeabilization, RNA fragmentation, 3'-end biotin labelling, proximity ligation, followed by RNA purification, enrichment of chimeric junctions, and strand-specific library preparation for sequencing. directional sequencing library was prepared using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB, #E7760), which employs a dUTP-based method with random hexamers for reverse transcription