Next-generation sequencing library construction on a surface

BMC Genomics
Kuan FengJeremy S Edwards

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

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Citations

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

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Datasets Mentioned

BETA
ERR481289

Methods Mentioned

BETA
PCR
Assay

Software Mentioned

samtools
BWA
Picardtools

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