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Closing gaps in the human genome with fosmid resources generated from multiple individuals

Abstract

The human genome sequence has been finished to very high standards; however, more than 340 gaps remained when the finished genome was published by the International Human Genome Sequencing Consortium in 2004. Using fosmid resources generated from multiple individuals, we targeted gaps in the euchromatic part of the human genome. Here we report 2,488,842 bp of previously unknown euchromatic sequence, 363,114 bp of which close 26 of 250 euchromatic gaps, or 10%, including two remaining euchromatic gaps on chromosome 19. Eight (30.7%) of the closed gaps were found to be polymorphic. These sequences allow complete annotation of several human genes as well as the assignment of mRNAs. The gap sequences are 2.3-fold enriched in segmentally duplicated sequences compared to the whole genome. Our analysis confirms that not all gaps within 'finished' genomes are recalcitrant to subcloning and suggests that the paired-end-sequenced fosmid libraries could prove to be a rich resource for completion of the human euchromatic genome.

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Figure 1: Strategies for closing or extending into gaps in the human genome.
Figure 2: Closing euchromatic gaps using haplotype-specific sequence assemblies.
Figure 3: Overview of CGH experiments for the closed gap-contigs.

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Acknowledgements

The authors would like to acknowledge the University of Washington Genome Center staff and A.Yamada at Agilent Technologies for technical assistance. This work was supported by US National Institutes of Health grants 3 U54 HG002043 to M.V.O. and HG004120 to E.E.E. and M.V.O. E.E.E. is an investigator of the Howard Hughes Medical Institute.

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Authors

Contributions

R.K. and M.V.O. designed the overall study. R.K. oversaw the overall data production and analysis. R.K. and E.E.E. carried out data analysis and wrote the manuscript with comments from M.V.O. D. Bovee was responsible for data curation, identification of clones from paired-end-sequenced fosmid libraries and incorporation of publicly available sequence data. E.H. and D.J. provided the informatics support and submission of the sequence data to Genbank. Y.Z. and Z.W. were responsible for finishing the fosmid clones. J.C. was responsible for custom fosmid library constructions, and R.L., D. Buckley and S.S. generated shotgun-sequencing data. H.S.H., W.G. and K.P. generated and analyzed the MCD fingerprint data. G.M.C. and N.S. designed and analyzed the aCGH data; E.E.E., E.T., V.A.M. and J.S. initially identified and cherry-picked the fosmid clones from G248 and ABC libraries. D.R.S. was responsible for generating ABC fosmid libraries and their paired-end-sequence data.

Corresponding author

Correspondence to Rajinder Kaul.

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Supplementary Tables 1–7 (PDF 560 kb)

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Bovee, D., Zhou, Y., Haugen, E. et al. Closing gaps in the human genome with fosmid resources generated from multiple individuals. Nat Genet 40, 96–101 (2008). https://doi.org/10.1038/ng.2007.34

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