Mycobacterium tuberculosis Erdman ATCC 35801 arrayed Himar1 transposon library isolated using MtbYM rich medium.

Abstract

We created a Himar1 transposon library in Mycobacterium tuberculosis Erdman ATCC 35801 strain containing ~8000 mutants arrayed in 80 tube racks in a 96-well format. The transposon library was isolated on MtbYM rich medium that allowed recovery of auxotrophic mutants that would not be viable on standard Middlebrook M. tuberculosis culture medium. Using orthogonal pooling and transposon sequencing (Tn-seq), we mapped the location of Tn mutants in the library. The library was split into two groups of 40 racks. For each set of 40 racks, pools were created of all mutants in each row (rows A-H), column (columns 1-12) and rack (racks 1-40 or 41-80) to generate 60 Tn-seq samples for Illumina sequencing. Two different mapping methods, the heuristic Straight Three strategy and the probabilistic Knockout Sudoku algorithm, were used to map the arrayed library with good agreement between both mapping methods. This repository contains Illumina sequencing data for all rack, column, and row pools used to map the entire 80 rack arrayed library.