title: Halobacterium salinarum NRC-1; Ura3 knockout; mid-exponential phase; biological replicate 2
design: Halobacterium salinarum knockout strains (delta-Ura3: control; delta-Ura3/delta-SmAP1: SmAP1 knockout) were cultured in uracil-supplemented (50 g/mL) complex medium (CM) until mid-exponential phase (OD600nm = 0.5). The cultures were grown at 37C, under light exposure, and constant agitation (125 RPM). We collected 2 mL samples and submitted them to DNA extraction using the DNeasy Blood & Tissue kit (QIAGEN), according to the manufacturer's instructions for Gram-negative bacteria. We tested DNA samples for purity and quantity using spectrophotometric and fluorimetric methods, respectively. The samples were prepared for long-read sequencing following the 1D native barcoding genomic DNA protocol using SQK-LSK108 and EXP-NBD103 (Oxford Nanopore Technologies). The equimolar pool of barcoded samples was sequenced using a MinION Mk1B instrument (Oxford Nanopore Technologies) in an FLO-MIN106 flow cell for 24 hours. Three biological replicates were sequenced for each one of the strains (control and SmAP1 knockout). Raw signal was demultiplexed using Deepbinner (0.1.1), base called using Guppy (2.3.7+e041753), and trimmed using Porechop (0.2.3). This sample corresponds to barcode 02 (control), biological replicate 2.