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MICROBIAL WHOLE GENOME SEQUENCING BEST PRACTICES With Single Molecule, Real-Time (SMRT®) Sequencing, you can affordably characterize complete microbial genomes. For most microbes, closed genomes and accessory plasmids can be assembled using PacBio data from single libraries in a single run — with turn-around times as short as one day. FROM GENOMIC DNA TO COMPLETE GENOME IN A SINGLE EXPERIMENT SAMPLE PREPARATION RECOMMENDATIONS -- Maximize throughput, efficiency, and project costs with -Ligate Barcode Adapters and Pool Samples for Multiplexing (Optional) BC1 Ligate Adapters for Single Sample microbial multiplexing1 -- Genomes with large repeats regions may require 20 kb or 30 kb library preparation with size selection2,3, and may not be suitable for multiplexing -- Complete genome assemblies may require reducing multiplex levels Use recommended starting input of high-quality DNA (1 µg) -- Low input options4, ranging from 10-100 ng are available Sequence to 50-fold coverage per microbe -- Sequel System: Multiplex up to 16 microbes (<2 Mb) or 12 microbes (<5 Mb) per SMRT Cell 1M -- PacBio RS II: Multiplex up to 2 microbes (<5 Mb) per SMRT Cell Automation solutions available GENERATE NEAR-COMPLETE MICROBIAL ASSEMBLIES WITH MULTIPLEXING AND SEQUEL SYSTEM Sample 16 SMRTbell™ adapters with barcodes Size Selection (Optional) Prepare Template(s) for Sequencing Sequence on PacBio Systems Sequel System Analyze with SMRT Analysis Software Suite A single Sequel SMRT Cell 1M can multiplex up to 16 microbes (<2 Mb) or 12 microbes (<5 Mb) and achieve near-complete assemblies (<10 contigs). Shown above are HGAP 4 de novo assembly statistics from an 8-plex pool of E. coli (5 Mb) strains and a 12-plex pool of B. subtilis (4 Mb) strains5. THE LEADER IN LONG-READ SEQUENC

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DATA ANALYSIS SOLUTIONS WITH SMRT® ANALYSIS AND PACBIO® DEVNET -- Generate exceptional genome and plasmid de novo assemblies with megabase-size contig N50s -- Achieve high-quality consensus accuracies > 99.999% -- Simultaneously characterize epigenetic modifications through kinetic analysis of base incorporation events during sequencing -- Easily demultiplex barcoded samples6, generate de novo assemblies7, and identify DNA modifications8 through a GUI-based user interface -- Utilize advanced data visualization and mining -- Output data in standard file formats - BAM and FASTA/Q - for...

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