Imperial College London

ProfessorPaulFreemont

Faculty of MedicineDepartment of Medicine

Chair in Protein Crystallography
 
 
 
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Contact

 

+44 (0)20 7594 5327p.freemont

 
 
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Location

 

259Sir Alexander Fleming BuildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Suckling:2019:10.1016/j.synbio.2019.01.002,
author = {Suckling, L and McFarlane, C and Sawyer, C and Chambers, SP and Kitney, RI and McClymont, DW and Freemont, PS},
doi = {10.1016/j.synbio.2019.01.002},
journal = {Synthetic and Systems Biotechnology},
pages = {57--66},
title = {Miniaturisation of high-throughput plasmid DNA library preparation for next-generation sequencing using multifactorial optimisation},
url = {http://dx.doi.org/10.1016/j.synbio.2019.01.002},
volume = {4},
year = {2019}
}

RIS format (EndNote, RefMan)

TY  - JOUR
AB - 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.
AU - Suckling,L
AU - McFarlane,C
AU - Sawyer,C
AU - Chambers,SP
AU - Kitney,RI
AU - McClymont,DW
AU - Freemont,PS
DO - 10.1016/j.synbio.2019.01.002
EP - 66
PY - 2019///
SN - 2405-805X
SP - 57
TI - Miniaturisation of high-throughput plasmid DNA library preparation for next-generation sequencing using multifactorial optimisation
T2 - Synthetic and Systems Biotechnology
UR - http://dx.doi.org/10.1016/j.synbio.2019.01.002
UR - http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000458661200008&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
UR - http://hdl.handle.net/10044/1/69621
VL - 4
ER -