Abstract
Molecular combing represents an advanced method for preparing extended DNA fibres. It enables uniform and parallel stretching of hundreds of DNA molecules on a glass surface, at an unprecedented resolution of two kilobase pairs per micrometre. When coupled with fluorescence in situ hybridization (FISH), molecular combing allows the direct visualisation of genomic structure and copy number variation, as well as the quantification of sizes of overlap and gap between sequence contigs in the genome assembly. Here we present a multicolor fibre-FISH protocol using DNA fibres prepared by molecular combing.
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Acknowledgements
We would like to thank Genomic Vision for kindly providing the protocols and for their constant availability and help in troubleshooting. We also thank our team members from the Molecular Cytogenetics Core Facility at the Wellcome Trust Sanger Institute, Beiyuan Fu and Ruby Banerjee as well as the former team member Elizabeth Langley, for the contributions in the optimisation of these protocols. The Molecular Cytogenetics Core Facility at the Wellcome Trust Sanger Institute is funded by the Wellcome Trust (grant number WT098051).
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Louzada, S., Komatsu, J., Yang, F. (2017). Fluorescence In Situ Hybridization onto DNA Fibres Generated Using Molecular Combing. In: Liehr, T. (eds) Fluorescence In Situ Hybridization (FISH). Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-52959-1_31
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DOI: https://doi.org/10.1007/978-3-662-52959-1_31
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Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-662-52957-7
Online ISBN: 978-3-662-52959-1
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