Visendi, Paul, Berkman, Paul J., Hayashi, Satomi, Golicz, Agnieszka A., Bayer, Philipp E., Ruperao, Pradeep, Hurgobin, Bhavna, Montenegro, Juan, Chan, Chon-Kit Kenneth, Staňková, Helena, Batley, Jacqueline, Šimková, Hana, Doležel, Jaroslav and Edwards, David (2016) An efficient approach to BAC based assembly of complex genomes. Plant Methods, 12 (2). pp. 1-9. ISSN 1746-4811 (Print), 1746-4811 (Online) (doi:10.1186/s13007-016-0107-9)

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Abstract

Background: There has been an exponential growth in the number of genome sequencing projects since the introduction of next generation DNA sequencing technologies. Genome projects have increasingly involved assembly of whole genome data which produces inferior assemblies compared to traditional Sanger sequencing of genomic fragments cloned into bacterial artificial chromosomes (BACs). While whole genome shotgun sequencing using next generation sequencing (NGS) is relatively fast and inexpensive, this method is extremely challenging for highly complex genomes, where polyploidy or high repeat content confounds accurate assembly, or where a highly accurate ‘gold’ reference is required. Several attempts have been made to improve genome sequencing approaches by incorporating NGS methods, to variable success.

Results: We present the application of a novel BAC sequencing approach which combines indexed pools of BACs, Illumina paired read sequencing, a sequence assembler specifically designed for complex BAC assembly, and a custom bioinformatics pipeline. We demonstrate this method by sequencing and assembling BAC cloned fragments from bread wheat and sugarcane genomes.

Conclusions: We demonstrate that our assembly approach is accurate, robust, cost effective and scalable, with applications for complete genome sequencing in large and complex genomes.

Item Type:	Article
Additional Information:	© 2016 Visendi et al. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Uncontrolled Keywords:	Next-generation sequencing – SASSY – BAC – Assembly – 7DS – Triticum aestivum – Saccharum spp
Faculty / School / Research Centre / Research Group:	Faculty of Engineering & Science > Natural Resources Institute > Molecular Virology and Entomology Research Group
Last Modified:	11 Apr 2017 08:50
URI:	http://gala.gre.ac.uk/id/eprint/14756

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