Reliable multiplex sequencing with rare index mis-assignment on DNB-based NGS platform

Massively-parallel-sequencing, coupled with sample multiplexing, has made genetic tests broadly affordable. However, intractable index mis-assignments (commonly exceeds 1%) were repeatedly reported on some widely used sequencing platforms. Here, we investigated this quality issue on BGI sequencers u...

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Published in:BMC genomics 2019-03, Vol.20 (1), p.215-215, Article 215
Main Authors: Li, Qiaoling, Zhao, Xia, Zhang, Wenwei, Wang, Lin, Wang, Jingjing, Xu, Dongyang, Mei, Zhiying, Liu, Qiang, Du, Shiyi, Li, Zhanqing, Liang, Xinming, Wang, Xiaman, Wei, Hanmin, Liu, Pengjuan, Zou, Jing, Shen, Hanjie, Chen, Ao, Drmanac, Snezana, Liu, Jia Sophie, Li, Li, Jiang, Hui, Zhang, Yongwei, Wang, Jian, Yang, Huanming, Xu, Xun, Drmanac, Radoje, Jiang, Yuan
Format: Article
Language:English
Subjects:
Deoxyribonucleic acid
DNA
DNA biosynthesis
DNA nanoball technology
DNA sequencing
Gene sequencing
Genetic research
Genetic testing
Genomes
Genomics
Libraries
Methods
Multiplex sequencing
Multiplexing
NGS
Nucleotide sequence
Polymerase chain reaction
Quality control
Rare index mis-assignment
Technology
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cited_by cdi_FETCH-LOGICAL-c594t-46b91c4e8de77122c04209cffb0c8fc28ea37bb1c2c27490d5998d9ffe3ceb1a3
cites cdi_FETCH-LOGICAL-c594t-46b91c4e8de77122c04209cffb0c8fc28ea37bb1c2c27490d5998d9ffe3ceb1a3
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container_title BMC genomics
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creator Li, Qiaoling
Zhao, Xia
Zhang, Wenwei
Wang, Lin
Wang, Jingjing
Xu, Dongyang
Mei, Zhiying
Liu, Qiang
Du, Shiyi
Li, Zhanqing
Liang, Xinming
Wang, Xiaman
Wei, Hanmin
Liu, Pengjuan
Zou, Jing
Shen, Hanjie
Chen, Ao
Drmanac, Snezana
Liu, Jia Sophie
Li, Li
Jiang, Hui
Zhang, Yongwei
Wang, Jian
Yang, Huanming
Xu, Xun
Drmanac, Radoje
Jiang, Yuan
description Massively-parallel-sequencing, coupled with sample multiplexing, has made genetic tests broadly affordable. However, intractable index mis-assignments (commonly exceeds 1%) were repeatedly reported on some widely used sequencing platforms. Here, we investigated this quality issue on BGI sequencers using three library preparation methods: whole genome sequencing (WGS) with PCR, PCR-free WGS, and two-step targeted PCR. BGI's sequencers utilize a unique DNA nanoball (DNB) technology which uses rolling circle replication for DNA-nanoball preparation; this linear amplification is PCR free and can avoid error accumulation. We demonstrated that single index mis-assignment from free indexed oligos occurs at a rate of one in 36 million reads, suggesting virtually no index hopping during DNB creation and arraying. Furthermore, the DNB-based NGS libraries have achieved an unprecedentedly low sample-to-sample mis-assignment rate of 0.0001 to 0.0004% under recommended procedures. Single indexing with DNB technology provides a simple but effective method for sensitive genetic assays with large sample numbers.
doi_str_mv 10.1186/s12864-019-5569-5
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subjects Deoxyribonucleic acid
DNA
DNA biosynthesis
DNA nanoball technology
DNA sequencing
Gene sequencing
Genetic research
Genetic testing
Genomes
Genomics
Libraries
Methods
Multiplex sequencing
Multiplexing
NGS
Nucleotide sequence
Polymerase chain reaction
Quality control
Rare index mis-assignment
Technology
title Reliable multiplex sequencing with rare index mis-assignment on DNB-based NGS platform
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