RT Journal
A1 Wang, Ou
A1 Chin, Robert
A1 Cheng, Xiaofang
A1 Wu, Michelle Ka Yan
A1 Mao, Qing
A1 Tang, Jingbo
A1 Sun, Yuhui
A1 Anderson, Ellis
A1 Lam, Han K.
A1 Chen, Dan
A1 Zhou, Yujun
A1 Wang, Linying
A1 Fan, Fei
A1 Zou, Yan
A1 Xie, Yinlong
A1 Zhang, Rebecca Yu
A1 Drmanac, Snezana
A1 Nguyen, Darlene
A1 Xu, Chongjun
A1 Villarosa, Christian
A1 Gablenz, Scott
A1 Barua, Nina
A1 Nguyen, Staci
A1 Tian, Wenlan
A1 Liu, Jia Sophie
A1 Wang, Jingwan
A1 Liu, Xiao
A1 Qi, Xiaojuan
A1 Chen, Ao
A1 Wang, He
A1 Dong, Yuliang
A1 Zhang, Wenwei
A1 Alexeev, Andrei
A1 Yang, Huanming
A1 Wang, Jian
A1 Kristiansen, Karsten
A1 Xu, Xun
A1 Drmanac, Radoje
A1 Peters, Brock A.
T1 Efficient and unique cobarcoding of second-generation sequencing reads from long DNA molecules enabling cost-effective and accurate sequencing, haplotyping, and de novo assembly
JF Genome Research 
JO Genome Research 
YR 2019 
FD May 01 
VO 29 
IS 5 
SP 798 
OP 808 
DO 10.1101/gr.245126.118 
UL http://genome.cshlp.org/content/29/5/798.abstract 
AB Here, we describe single-tube long fragment read (stLFR), a technology that enables sequencing of data from long DNA molecules using economical second-generation sequencing technology. It is based on adding the same barcode sequence to subfragments of the original long DNA molecule (DNA cobarcoding). To achieve this efficiently, stLFR uses the surface of microbeads to create millions of miniaturized barcoding reactions in a single tube. Using a combinatorial process, up to 3.6 billion unique barcode sequences were generated on beads, enabling practically nonredundant cobarcoding with 50 million barcodes per sample. Using stLFR, we demonstrate efficient unique cobarcoding of more than 8 million 20- to 300-kb genomic DNA fragments. Analysis of the human genome NA12878 with stLFR demonstrated high-quality variant calling and phase block lengths up to N50 34 Mb. We also demonstrate detection of complex structural variants and complete diploid de novo assembly of NA12878. These analyses were all performed using single stLFR libraries, and their construction did not significantly add to the time or cost of whole-genome sequencing (WGS) library preparation. stLFR represents an easily automatable solution that enables high-quality sequencing, phasing, SV detection, scaffolding, cost-effective diploid de novo genome assembly, and other long DNA sequencing applications.