single-tube LFR

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.单管长片段读取（single tube long fragment read, stLFR）技术可实现高效的全基因组测序（Whole Genome Sequencing, WGS）、单倍型分型以及重叠群支架构建。该技术的核心原理是为原始DNA分子的亚片段添加相同的条形码序列（DNA共条形码标记，DNA co-barcoding）。为达成这一目标，stLFR技术借助微珠表面在单管体系中构建出数百万个微型化反应隔间。通过组合式合成流程，微珠表面可生成超过18亿条独特的条形码序列，使得在包含5000万条条形码的反应体系中可实现近乎无冗余的共条形码标记。利用stLFR技术，本研究实现了对超过800万条20~300 kb的基因组DNA片段的高效特异性共条形码标记，并对NA12878基因组完成了近乎完美的变异检测与单倍型分型，将其重叠群N50提升至23.4 Mb。stLFR是一种低成本的单文库解决方案，可助力获取长读长序列数据。

搜集汇总

背景与挑战

背景概述

该数据集基于单管长片段读取（stLFR）技术，通过微珠表面创建数百万个微型隔室，实现高效的全基因组测序、单倍型分型和contig支架构建。数据包含人类基因组NA12878的测序数据，展示了近完美的变异检测和单倍型分型能力，N50达到23.4 Mb。数据集由14个样品、28个实验组成，总数据量4.96TB。