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Miniaturisation of high-throughput plasmid DNA library preparation for next-generation sequencing using multifactorial optimisation

Title: Miniaturisation of high-throughput plasmid DNA library preparation for next-generation sequencing using multifactorial optimisation
Authors: Suckling, L
McFarlane, C
Sawyer, C
Chambers, SP
Kitney, RI
McClymont, DW
Freemont, PS
Item Type: Journal Article
Abstract: High-throughput preparation of plasmid DNA libraries for next-generation sequencing (NGS) is an important capability for molecular biology laboratories. In particular, it is an essential quality control (QC) check when large numbers of plasmid variants are being generated. Here, we describe the use of the Design of Experiments (DOE) methodology to optimise the miniaturised preparation of plasmid DNA libraries for NGS, using the Illumina® Nextera XT technology and the Labcyte Echo® acoustic liquid dispensing system. Furthermore, we describe methods which can be implemented as a QC check for identifying the presence of genomic DNA (gDNA) in plasmid DNA samples and the subsequent shearing of the gDNA, which otherwise prevents the acoustic transfer of plasmid DNA. This workflow enables the preparation of plasmid DNA libraries which yield high-quality sequencing data.
Issue Date: 1-Mar-2019
Date of Acceptance: 16-Jan-2019
URI: http://hdl.handle.net/10044/1/69621
DOI: https://dx.doi.org/10.1016/j.synbio.2019.01.002
ISSN: 2405-805X
Publisher: KeAi
Start Page: 57
End Page: 66
Journal / Book Title: Synthetic and Systems Biotechnology
Volume: 4
Issue: 1
Copyright Statement: © 2019 Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/BY-NC-ND/4.0/).
Sponsor/Funder: Engineering & Physical Science Research Council (EPSRC)
Engineering & Physical Science Research Council (EPSRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
Engineering & Physical Science Research Council (EPSRC)
Funder's Grant Number: EP/L011573/1
EP/M006700/1
BB/M025632/1
EP/N023854/1
Keywords: Science & Technology
Life Sciences & Biomedicine
Biotechnology & Applied Microbiology
Information & Library Sciences
0807 Library and Information Studies
Publication Status: Published
Online Publication Date: 2019-01-22
Appears in Collections:Bioengineering
Department of Medicine
Faculty of Medicine



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