Oxford Nanopore genome sequencing of a virulent barcodedM. tuberculosis Erdman strain
An aliquot of the frozen barcoded Erdman library used for NHP infections (NR-50781, BEI resources) was plated on 7H10 plates at 37C. A single colony was grown in 7H9 broth with OADC supplement at 37C and bacterial genomic DNA extracted using phenol-chloroform extract6ion (see methods in Github) and sent for commercial sequencing using paired long read Nanopore and short-read Illumina sequencing (SeqCenter, Pittsburgh). The library was prepared using Oxford Nanopore Technologies (ONT) Native Barcoding Kit 24 V14 (SQK-NBD114.24) and run on a R10.4.1 flow cell. Samples were demultiplexed using Guppys Super Accuratebasecalling model (dna_r10.4.1_e8.2_400bps_modbases_5mc_cg_sup.cfg).