Efficient long single molecule sequencing for cost effective and accurate sequencing, haplotyping, and de novo assembly
Michelle Ka Wu,
Han K. Lam,
Rebecca Yu Zhang,
Jia Sophie Liu,
Brock A. Peters
Posted 17 May 2018
bioRxiv DOI: 10.1101/324392 (published DOI: 10.1101/gr.245126.118)
Posted 17 May 2018
Single tube long fragment read (stLFR) technology enables efficient WGS, haplotyping, and contig scaffolding. It is based on adding the same barcode sequence to sub-fragments of the original DNA molecule (DNA co-barcoding). To achieve this, stLFR uses the surface of microbeads to create millions of miniaturized compartments in a single tube. Using a combinatorial process over 1.8 billion unique barcode sequences were generated on beads, enabling practically non-redundant co-barcoding in reactions with 50 million barcodes. Using stLFR we demonstrate efficient unique co-barcoding of over 8 million 20-300 kb genomic DNA fragments with near perfect variant calling and phasing of the genome of NA12878 into contigs up to N50 23.4 Mb. stLFR represents a low-cost single library solution that can enable long sequence data.
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