Comparative genomic hybridization using oligonucleotide microarrays and total genomic DNA
| العنوان: | Comparative genomic hybridization using oligonucleotide microarrays and total genomic DNA |
|---|---|
| المؤلفون: | Michael T. Barrett, Robert Kincaid, Paul S. Meltzer, Alicia F. Scheffer, Kristin Baird, Nick Sampas, Doron Lipson, Zohar Yakhini, Bo Curry, Stephen Laderman, Laurakay Bruhn, Peter Tsang, Amir Ben-Dor |
| المصدر: | Proceedings of the National Academy of Sciences. 101:17765-17770 |
| بيانات النشر: | Proceedings of the National Academy of Sciences, 2004. |
| سنة النشر: | 2004 |
| مصطلحات موضوعية: | Male, Biology, Sensitivity and Specificity, Genome, Cell Line, Complementary DNA, Humans, X chromosome, Oligonucleotide Array Sequence Analysis, Chromosome Aberrations, Genetics, Multidisciplinary, Gene Expression Profiling, Homozygote, Nucleic Acid Hybridization, DNA, Biological Sciences, Amplicon, genomic DNA, Research Design, Female, DNA microarray, Gene Deletion, Virtual karyotype, Comparative genomic hybridization |
| الوصف: | Array-based comparative genomic hybridization (CGH) measures copy-number variations at multiple loci simultaneously, providing an important tool for studying cancer and developmental disorders and for developing diagnostic and therapeutic targets. Arrays for CGH based on PCR products representing assemblies of BAC or cDNA clones typically require maintenance, propagation, replication, and verification of large clone sets. Furthermore, it is difficult to control the specificity of the hybridization to the complex sequences that are present in each feature of such arrays. To develop a more robust and flexible platform, we created probe-design methods and assay protocols that make oligonucleotide microarrays synthesized in situ by inkjet technology compatible with array-based comparative genomic hybridization applications employing samples of total genomic DNA. Hybridization of a series of cell lines with variable numbers of X chromosomes to arrays designed for CGH measurements gave median ratios for X-chromosome probes within 6% of the theoretical values (0.5 for XY/XX, 1.0 for XX/XX, 1.4 for XXX/XX, 2.1 for XXXX/XX, and 2.6 for XXXXX/XX). Furthermore, these arrays detected and mapped regions of single-copy losses, homozygous deletions, and amplicons of various sizes in different model systems, including diploid cells with a chromosomal breakpoint that has been mapped and sequenced to a precise nucleotide and tumor cell lines with highly variable regions of gains and losses. Our results demonstrate that oligonucleotide arrays designed for CGH provide a robust and precise platform for detecting chromosomal alterations throughout a genome with high sensitivity even when using full-complexity genomic samples. |
| تدمد: | 1091-6490 0027-8424 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::69d6482ace21381788745201a3385d1c https://doi.org/10.1073/pnas.0407979101 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....69d6482ace21381788745201a3385d1c |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10916490 00278424 |
|---|