• Sequence specificity of streptozotocin-induced mutations

      Mack, Susan L.; Fram, Robert J.; Marinus, Martin G. (1988-10-25)
      The isolation and characterization of streptozotocin (STZ)-induced mutations in the phage P22 mnt repressor gene is described. Cells carrying the plasmid-borne mnt gene were exposed to STZ to give 10-20 percent survival and at least an eleven-fold increase in mutation frequency. DNA sequence analysis showed that 50 of 51 STZ-induced mutations were GC to AT transitions, and one was an AT to GC transition. We have also compared the STZ mutational spectrum to that for N-methyl-N'-nitro-N-nitroso-guanidine (MNNG). There are sites in the mnt gene which are mutated only by STZ; only by MNNG, or by both agents. Sites at which only STZ induced GC to AT transition mutations occur were in sequences that are pyrimidine rich 5' to the mutated site and purine rich 3' to the mutated site. Induction of mutations by both STZ and MNNG should be considered to maximize the number of mutable sites.
    • Simultaneous generation of many RNA-seq libraries in a single reaction

      Shishkin, Alexander A.; Giannoukos, Georgia; Kucukural, Alper; Ciulla, Dawn; Busby, Michele; Surka, Christine; Chen, Jenny; Bhattacharyya, Roby P.; Rudy, Robert F.; Patel, Milesh M.; et al. (2015-04-01)
      Although RNA-seq is a powerful tool, the considerable time and cost associated with library construction has limited its utilization for various applications. RNAtag-Seq, an approach to generate multiple RNA-seq libraries in a single reaction, lowers time and cost per sample, and it produces data on prokaryotic and eukaryotic samples that are comparable to those generated by traditional strand-specific RNA-seq approaches.