Phage tRNA library screening in E. coli: post-activation of AriB, replicate 1
Plasmids were isolated from E. coli using the Monarch Plasmid Miniprep Kit (NEB, T1010S). To amplify the phage tRNA sequences encoded on them, the plasmids were subjected to PCR using Q5 polymerase and primers containing the p5 and p7 adaptor sequences. Once amplified, the tRNA sequences were amplified once more with Q5 and primers with the Illumina i5 and i7 index adaptor sequences. Once purified via magnetic bead purification, amplicons were diluted in 10 mM Tris/HCl pH 8 and loaded into the iSeq 100 System using the 300-cycle iSeq 100 i1 Reagent v2 for single-end sequencing.