Abstract
AbstractWe have designed DOP‐PCR primers specifically for the amplification of large insert clones for use in the construction of DNA microarrays. A bioinformatic approach was used to construct primers that were efficient in the general amplification of human DNA but were poor at amplifying E. coli DNA, a common contaminant of DNA preparations from large insert clones. We chose the three most selective primers for use in printing DNA microarrays. DNA combined from the amplification of large insert clones by use of these three primers and spotted onto glass slides showed more than a sixfold increase in the human to E. coli hybridization ratio when compared to the standard DOP‐PCR primer, 6MW. The microarrays reproducibly delineated previously characterized gains and deletions in a cancer cell line and identified a small gain not detected by use of conventional CGH. We also describe a method for the bulk testing of the hybridization characteristics of chromosome‐specific clones spotted on microarrays by use of DNA amplified from flow‐sorted chromosomes. Finally, we describe a set of clones selected from the publicly available Golden Path of the human genome at 1‐Mb intervals and a view in the Ensembl genome browser from which data required for the use of these clones in array CGH and other experiments can be downloaded across the Internet. © 2003 Wiley‐Liss, Inc.
Bibliography
Fiegler, H., Carr, P., Douglas, E. J., Burford, D. C., Hunt, S., Smith, J., Vetrie, D., Gorman, P., Tomlinson, I. P. M., & Carter, N. P. (2003). DNA microarrays for comparative genomic hybridization based on DOPâPCR amplification of BAC and PAC clones. Genes, Chromosomes and Cancer, 36(4), 361â374. Portico.
References
18
Referenced
353
{'key': 'e_1_2_6_2_1', 'first-page': '187', 'article-title': 'Bivariate chromosome analysis using a commercial flow cytometer', 'volume': '29', 'author': 'Carter NP', 'year': '1994', 'journal-title': 'Methods Mol Biol'}
/ Methods Mol Biol / Bivariate chromosome analysis using a commercial flow cytometer by Carter NP (1994)10.2144/00293bm01
/ Biotechniques / Real‐time PCR‐based method for assaying the purity of bacterial artificial chromosome preparations by Foreman PK (2000)10.1038/ng771
{'key': 'e_1_2_6_5_1', 'first-page': '69', 'article-title': 'Contiguation of bacterial clones', 'volume': '175', 'author': 'Humphray SJ', 'year': '2001', 'journal-title': 'Methods Mol Biol'}
/ Methods Mol Biol / Contiguation of bacterial clones by Humphray SJ (2001)10.1126/science.1359641
10.1038/35057062
10.1016/S1525-1578(10)60634-4
10.1016/S0037-1963(00)90015-5
10.1101/gr.7.11.1072
10.1038/2524
10.1038/ng0495-369
{'key': 'e_1_2_6_13_1', 'volume-title': 'Genome mapping: a practical approach', 'author': 'Ross MT', 'year': '1997'}
/ Genome mapping: a practical approach by Ross MT (1997){'key': 'e_1_2_6_14_1', 'first-page': '1433', 'article-title': 'Comparative genomic hybridization of breast tumors stratified by histological grade reveals new insights into the biological progression of breast cancer', 'volume': '59', 'author': 'Roylance R', 'year': '1999', 'journal-title': 'Cancer Res'}
/ Cancer Res / Comparative genomic hybridization of breast tumors stratified by histological grade reveals new insights into the biological progression of breast cancer by Roylance R (1999)10.1038/ng754
10.1002/(SICI)1098-2264(199712)20:4<399::AID-GCC12>3.0.CO;2-I
10.1016/0888-7543(92)90147-K
10.1002/gcc.2870040311
10.1086/340426
Dates
Type | When |
---|---|
Created | 22 years, 5 months ago (March 18, 2003, 5:54 p.m.) |
Deposited | 1 year, 9 months ago (Nov. 17, 2023, 9:32 a.m.) |
Indexed | 1 month, 3 weeks ago (July 12, 2025, 6:54 p.m.) |
Issued | 22 years, 7 months ago (Jan. 30, 2003) |
Published | 22 years, 7 months ago (Jan. 30, 2003) |
Published Online | 22 years, 7 months ago (Jan. 30, 2003) |
Published Print | 22 years, 5 months ago (April 1, 2003) |
@article{Fiegler_2003, title={DNA microarrays for comparative genomic hybridization based on DOP‐PCR amplification of BAC and PAC clones}, volume={36}, ISSN={1098-2264}, url={http://dx.doi.org/10.1002/gcc.10155}, DOI={10.1002/gcc.10155}, number={4}, journal={Genes, Chromosomes and Cancer}, publisher={Wiley}, author={Fiegler, Heike and Carr, Philippa and Douglas, Eleanor J. and Burford, Deborah C. and Hunt, Sarah and Smith, James and Vetrie, David and Gorman, Patricia and Tomlinson, Ian P.M. and Carter, Nigel P.}, year={2003}, month=jan, pages={361–374} }