Ultralow-input single-tube linked-read library method enables short-read second-generation sequencing systems to routinely generate highly accurate and economical long-range sequencing information [METHOD]
Long-range sequencing information is required for haplotype phasing, de novo assembly, and structural variation detection. Current long-read sequencing technologies can provide valuable long-range information but at a high cost with low accuracy and high DNA input requirements. We have developed a single-tube Transposase Enzyme Linked Long-read Sequencing (TELL-seq) technology, which enables a low-cost, high-accuracy, and high-throughput short-read second-generation sequencer to generate over 100 kb of long-range sequencing information with as little as 0.1 ng input material. In a PCR tube, millions of clonally barcoded beads are used to uniquely barcode long DNA molecules in an open bulk reaction without dilution and compartmentation. The barcoded linked-reads are used to successfully assemble genomes ranging from microbes to human. These linked-reads also generate megabase-long phased blocks and provide a cost-effective tool for detecting structural variants in a genome, which are important to identify compound heterozygosity in recessive Mendelian diseases and discover genetic drivers and diagnostic biomarkers in cancers.
Source: Genome Research - Category: Genetics & Stem Cells Authors: Chen, Z., Pham, L., Wu, T.-C., Mo, G., Xia, Y., Chang, P. L., Porter, D., Phan, T., Che, H., Tran, H., Bansal, V., Shaffer, J., Belda-Ferre, P., Humphrey, G., Knight, R., Pevzner, P., Pham, S., Wang, Y., Lei, M. Tags: METHOD Source Type: research