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Human and mouse oligonucleotide-based array CGH
Array-based comparative genomic hybridization is a high resolution method for measuring chromosomal copy number changes. Here we present a validated protocol using in-house spotted oligonucleotide libraries for array comparative genomic hybridization (CGH). This oligo array CGH platform yields repro...
Autores principales: | , , , , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2005
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1316119/ https://www.ncbi.nlm.nih.gov/pubmed/16361265 http://dx.doi.org/10.1093/nar/gni191 |
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author | van den IJssel, Paul Tijssen, Marianne Chin, Suet-Feung Eijk, Paul Carvalho, Beatriz Hopmans, Erik Holstege, Henne Bangarusamy, Dhinoth Kumar Jonkers, Jos Meijer, Gerrit A. Caldas, Carlos Ylstra, Bauke |
author_facet | van den IJssel, Paul Tijssen, Marianne Chin, Suet-Feung Eijk, Paul Carvalho, Beatriz Hopmans, Erik Holstege, Henne Bangarusamy, Dhinoth Kumar Jonkers, Jos Meijer, Gerrit A. Caldas, Carlos Ylstra, Bauke |
author_sort | van den IJssel, Paul |
collection | PubMed |
description | Array-based comparative genomic hybridization is a high resolution method for measuring chromosomal copy number changes. Here we present a validated protocol using in-house spotted oligonucleotide libraries for array comparative genomic hybridization (CGH). This oligo array CGH platform yields reproducible results and is capable of detecting single copy gains, multi-copy amplifications as well as homozygous and heterozygous deletions as small as 100 kb with high resolution. A human oligonucleotide library was printed on amine binding slides. Arrays were hybridized using a hybstation and analysed using BlueFuse feature extraction software, with >95% of spots passing quality control. The protocol allows as little as 300 ng of input DNA and a 90% reduction of Cot-1 DNA without compromising quality. High quality results have also been obtained with DNA from archival tissue. Finally, in addition to human oligo arrays, we have applied the protocol successfully to mouse oligo arrays. We believe that this oligo-based platform using ‘off-the-shelf’ oligo libraries provides an easy accessible alternative to BAC arrays for CGH, which is cost-effective, available at high resolution and easily implemented for any sequenced organism without compromising the quality of the results. |
format | Text |
id | pubmed-1316119 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2005 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-13161192005-12-19 Human and mouse oligonucleotide-based array CGH van den IJssel, Paul Tijssen, Marianne Chin, Suet-Feung Eijk, Paul Carvalho, Beatriz Hopmans, Erik Holstege, Henne Bangarusamy, Dhinoth Kumar Jonkers, Jos Meijer, Gerrit A. Caldas, Carlos Ylstra, Bauke Nucleic Acids Res Methods Online Array-based comparative genomic hybridization is a high resolution method for measuring chromosomal copy number changes. Here we present a validated protocol using in-house spotted oligonucleotide libraries for array comparative genomic hybridization (CGH). This oligo array CGH platform yields reproducible results and is capable of detecting single copy gains, multi-copy amplifications as well as homozygous and heterozygous deletions as small as 100 kb with high resolution. A human oligonucleotide library was printed on amine binding slides. Arrays were hybridized using a hybstation and analysed using BlueFuse feature extraction software, with >95% of spots passing quality control. The protocol allows as little as 300 ng of input DNA and a 90% reduction of Cot-1 DNA without compromising quality. High quality results have also been obtained with DNA from archival tissue. Finally, in addition to human oligo arrays, we have applied the protocol successfully to mouse oligo arrays. We believe that this oligo-based platform using ‘off-the-shelf’ oligo libraries provides an easy accessible alternative to BAC arrays for CGH, which is cost-effective, available at high resolution and easily implemented for any sequenced organism without compromising the quality of the results. Oxford University Press 2005 2005-12-16 /pmc/articles/PMC1316119/ /pubmed/16361265 http://dx.doi.org/10.1093/nar/gni191 Text en © The Author 2005. Published by Oxford University Press. All rights reserved |
spellingShingle | Methods Online van den IJssel, Paul Tijssen, Marianne Chin, Suet-Feung Eijk, Paul Carvalho, Beatriz Hopmans, Erik Holstege, Henne Bangarusamy, Dhinoth Kumar Jonkers, Jos Meijer, Gerrit A. Caldas, Carlos Ylstra, Bauke Human and mouse oligonucleotide-based array CGH |
title | Human and mouse oligonucleotide-based array CGH |
title_full | Human and mouse oligonucleotide-based array CGH |
title_fullStr | Human and mouse oligonucleotide-based array CGH |
title_full_unstemmed | Human and mouse oligonucleotide-based array CGH |
title_short | Human and mouse oligonucleotide-based array CGH |
title_sort | human and mouse oligonucleotide-based array cgh |
topic | Methods Online |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1316119/ https://www.ncbi.nlm.nih.gov/pubmed/16361265 http://dx.doi.org/10.1093/nar/gni191 |
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