Rappaport Israeli genome
DNA was extracted from 2 ml of culture using the DNeasy Blood and Tissue Kit (Qiagen) as per manufacturer’s instructions. For each sample 100 ?g of DNA was used to prepare a barcoded library for sequencing, using the Ion Xpress™ Plus Fragment Library Kit (Life Technologies) per manufacturer’s instructions. In short, DNA was fragmented with Ion Shear Enzyme Mix II, and adaptors and barcodes were ligated to the fragmented DNA. The ligated DNA was then size-selected on a 2% E-Gel® SizeSelect™ Agarose Gel (Invitrogen). The size-selected library was then amplified using the primers supplied in the Ion Xpress™ Plus Fragment Library Kit (Life Technologies) per manufacturer’s instructions. The concentrations of the libraries were measured using a Qubit® dsDNA HS Assay Kit, and the size distribution was analyzed with an Agilent® High Sensitivity DNA Kit. Libraries were prepared for sequencing on a One Touch 2 using the Ion PGM™ Template OT2 200 Kit (Life Technologies) as per manufacturer’s instructions. The sequencing was then performed on an Ion Torrent PGM using Ion PGM™ Sequencing 200 Kit v2 (Life Technologies).