Nanopore small gDNA sequencing (long reads)

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The native barcoding kit enhances nanopore sequencing by enabling simultaneous sequencing of multiple samples with unique barcodes. After gDNA extraction, each sample is tagged with a barcode during library preparation. Sequencing generates long reads, allowing real-time base calling. Bioinformatics tools can be used to analyze data, including aligning reads and variant calling. This approach is particularly useful for studying small bacteria genomes because it is cost-effective and efficient, beneficial for microbiome studies and environmental monitoring and other applications in microbial genomics.

Libraries are prepared with the Native Barcoding Kit 96 V14 that enables PCR-free multiplexing of small gDNA samples using 96 unique barcodes. It involves repairing and dA-tailing gDNA, then ligating a unique dT-tailed barcode adapter. Barcoded samples are pooled, and each barcode adapter ligates to a sequencing adapter. Optimized for high sequencing accuracies (>99% Q20+) on nanopore Flowcells R10.4.1.