| description |
This experiment consists of two studies. The first where two independent biological replicates of MG1655, MG1655 thyA suhB::thyA and MG1655 thyA nusB::thyA were each grown in LB to mid-exponential phase. RNA-seq and associated data analysis were performed as described previously (Stringer et al., 2014. PMID 24272778). The second where five cultures of MG1655 thyA suhB::thyA were grown overnight from single colonies at 37 C in LB. 5 L of each overnight culture was spread on LB agar and incubated at 30 C, the non-permissive temperature for suhB mutants. One suppressor mutant colony was selected from each plate. rnc was PCR amplified from colonies using oligonucleotides JW836-JW837, and the PCR products were sequenced to identify the presence, if any, of suppressor mutations. Genomic DNA from a strain with wild-type rnc was prepared using a DNeasy Blood and Tissue Kit (Qiagen). A DNA library was prepared using a Nextera kit (Illumina). The library was sequenced (paired-end reads) using an Illumina MiSeq instrument. |