Validation of DNA probes for molecular cytogenetics by mapping onto immobilized circular DNA
<p>Abstract</p> <p>Background</p> <p>Fluorescence <it>in situ </it>hybridization (FISH) is a sensitive and rapid procedure to detect gene rearrangements in tumor cells using non-isotopically labeled DNA probes. Large insert recombinant DNA clones such as bac...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2008-12-01
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| Series: | Molecular Cytogenetics |
| Online Access: | http://www.molecularcytogenetics.org/content/1/1/28 |
| _version_ | 1818915928828542976 |
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| author | Rhein Andreas P Wang Mei Greulich-Bode Karin M Weier Jingly F Weier Heinz-Ulli G |
| author_facet | Rhein Andreas P Wang Mei Greulich-Bode Karin M Weier Jingly F Weier Heinz-Ulli G |
| author_sort | Rhein Andreas P |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Fluorescence <it>in situ </it>hybridization (FISH) is a sensitive and rapid procedure to detect gene rearrangements in tumor cells using non-isotopically labeled DNA probes. Large insert recombinant DNA clones such as bacterial artificial chromosome (BAC) or P1/PAC clones have established themselves in recent years as preferred starting material for probe preparations due to their low rates of chimerism and ease of use. However, when developing probes for the quantitative analysis of rearrangements involving genomic intervals of less than 100 kb, careful probe selection and characterization are of paramount importance.</p> <p>Results</p> <p>We describe a sensitive approach to quality control probe clones suspected of carrying deletions or for measuring clone overlap with near kilobase resolution. The method takes advantage of the fact that P1/PAC/BAC's can be isolated as circular DNA molecules, stretched out on glass slides and fine-mapped by multicolor hybridization with smaller probe molecules. Two examples demonstrate the application of this technique: mapping of a gene-specific ~6 kb plasmid onto an unusually small, ~55 kb circular P1 molecule and the determination of the extent of overlap between P1 molecules homologous to the human NF-κB2 locus.</p> <p>Conclusion</p> <p>The relatively simple method presented here does not require specialized equipment and may thus find widespread applications in DNA probe preparation and characterization, the assembly of physical maps for model organisms or in studies on gene rearrangements.</p> |
| first_indexed | 2024-12-20T00:10:05Z |
| format | Article |
| id | doaj.art-c9c866f62cde49619adcc1caa17c225d |
| institution | Directory Open Access Journal |
| issn | 1755-8166 |
| language | English |
| last_indexed | 2024-12-20T00:10:05Z |
| publishDate | 2008-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Molecular Cytogenetics |
| spelling | doaj.art-c9c866f62cde49619adcc1caa17c225d2022-12-21T20:00:32ZengBMCMolecular Cytogenetics1755-81662008-12-01112810.1186/1755-8166-1-28Validation of DNA probes for molecular cytogenetics by mapping onto immobilized circular DNARhein Andreas PWang MeiGreulich-Bode Karin MWeier Jingly FWeier Heinz-Ulli G<p>Abstract</p> <p>Background</p> <p>Fluorescence <it>in situ </it>hybridization (FISH) is a sensitive and rapid procedure to detect gene rearrangements in tumor cells using non-isotopically labeled DNA probes. Large insert recombinant DNA clones such as bacterial artificial chromosome (BAC) or P1/PAC clones have established themselves in recent years as preferred starting material for probe preparations due to their low rates of chimerism and ease of use. However, when developing probes for the quantitative analysis of rearrangements involving genomic intervals of less than 100 kb, careful probe selection and characterization are of paramount importance.</p> <p>Results</p> <p>We describe a sensitive approach to quality control probe clones suspected of carrying deletions or for measuring clone overlap with near kilobase resolution. The method takes advantage of the fact that P1/PAC/BAC's can be isolated as circular DNA molecules, stretched out on glass slides and fine-mapped by multicolor hybridization with smaller probe molecules. Two examples demonstrate the application of this technique: mapping of a gene-specific ~6 kb plasmid onto an unusually small, ~55 kb circular P1 molecule and the determination of the extent of overlap between P1 molecules homologous to the human NF-κB2 locus.</p> <p>Conclusion</p> <p>The relatively simple method presented here does not require specialized equipment and may thus find widespread applications in DNA probe preparation and characterization, the assembly of physical maps for model organisms or in studies on gene rearrangements.</p>http://www.molecularcytogenetics.org/content/1/1/28 |
| spellingShingle | Rhein Andreas P Wang Mei Greulich-Bode Karin M Weier Jingly F Weier Heinz-Ulli G Validation of DNA probes for molecular cytogenetics by mapping onto immobilized circular DNA Molecular Cytogenetics |
| title | Validation of DNA probes for molecular cytogenetics by mapping onto immobilized circular DNA |
| title_full | Validation of DNA probes for molecular cytogenetics by mapping onto immobilized circular DNA |
| title_fullStr | Validation of DNA probes for molecular cytogenetics by mapping onto immobilized circular DNA |
| title_full_unstemmed | Validation of DNA probes for molecular cytogenetics by mapping onto immobilized circular DNA |
| title_short | Validation of DNA probes for molecular cytogenetics by mapping onto immobilized circular DNA |
| title_sort | validation of dna probes for molecular cytogenetics by mapping onto immobilized circular dna |
| url | http://www.molecularcytogenetics.org/content/1/1/28 |
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