Abstract
Next-generation sequencing (NGS) has revolutionized almost all fields of biology, agriculture and medicine, and is widely utilized to analyse genetic variation. Over the past decade, the NGS pipeline has been steadily improved, and the entire process is currently relatively straightforward. However, NGS instrumentation still requires upfront library preparation, which can be a laborious process, requiring significant hands-on time. Herein, we present a simple but robust approach to streamline library preparation by utilizing surface bound transposases to construct DNA libraries directly on a flowcell surface. The surface bound transposases directly fragment genomic DNA while simultaneously attaching the library molecules to the flowcell. We sequenced and analysed a Drosophila genome library generated by this surface tagmentation approach, and we showed that our surface bound library quality was comparable to the quality of the library from a commercial kit. In addition to the time and cost savings, our approach does not require PCR amplification of the library, which eliminates potential problems associated with PCR duplicates. We described the first study to construct libraries directly on a flowcell. We believe our technique ...Continue Reading
References
Sep 30, 1994·Science·A BensimonD Bensimon
Nov 26, 1999·Nucleic Acids Research·R D Mitra, G M Church
Mar 25, 2000·Science·M D AdamsJ C Venter
Jul 31, 2003·Nucleic Acids Research·Joshua MerrittJeremy S Edwards
Mar 26, 2004·BMC Genetics·James A ButzJeremy S Edwards
Mar 31, 2004·Biotechnology and Bioengineering·Venugopal MikkilineniJeremy S Edwards
Sep 30, 2005·Biotechnology and Bioengineering·Joshua MerrittJeremy S Edwards
May 24, 2006·Nucleic Acids Research·Mindy SteinigerWilliam S Reznikoff
Nov 7, 2008·Nature·David R BentleyAnthony J Smith
May 20, 2009·Bioinformatics·Heng Li, Richard Durbin
Jun 10, 2009·Bioinformatics·Heng LiUNKNOWN 1000 Genome Project Data Processing Subgroup
Dec 15, 2010·Genome Biology·Andrew AdeyJay Shendure
Dec 15, 2010·Nanotechnology·Annegret BenkeWolfgang Pompe
Mar 25, 2011·Methods in Molecular Biology·Nicholas Caruccio
Nov 1, 2012·Proceedings of the National Academy of Sciences of the United States of America·Jerrod J SchwartzJay Shendure
Feb 8, 2014·BioTechniques·Steven R HeadPhillip Ordoukhanian
Aug 1, 2014·Genome Research·Simone PicelliRickard Sandberg
Aug 12, 2014·Trends in Genetics : TIG·Erwin L van DijkClaude Thermes
Feb 26, 2016·The AAPS Journal·Robert N Schuck, Joseph A Grillo
Citations
Jun 14, 2018·Briefings in Bioinformatics·Jiangning SongKuo-Chen Chou
Jul 22, 2019·Molecular Ecology Resources·Dandan LangXin Zhou
Jun 15, 2021·Frontiers in Oncology·Cristian ScatenaSara Tomei
Aug 8, 2019·ACS Applied Materials & Interfaces·Justin A CostaWei Zhou
Jun 18, 2020·ACS Omega·Xinyao MiaoBao Zhang