title: NextSeq 500 paired end sequencing; Ribo-seq in Escherichia coli MG1655 treated withTetracycline and Tylosin
design: Ribo-seq in Escherichia coli MG1655 treated withTetracycline and Tylosin
library_construction_protocol: Cultures were filtered at 17 min post-treatment. 200 ml of cells were grown in LB medium until OD600~1.1. One set of cultures was left untreated, other sets were treated with 1 μg/ml tetracycline or 400 μg/ml tylosin. Monosomes were isolated as previously described (Smith et al 2019 biorXiv) Library was constructed as previously described (Smith et al 2019 biorXiv) with minor modification: 3' linked oligonucleotide ligation reaction was carried overnight at 16oC. Circularized, reverse-transcribed cDNA was PCR-amplified for 6-8 cycles using primers containing Illumina sequencing indices