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| HIGH-RESOLUTION DNA FIBER-FISH FOR GENOMIC DNA MAPPING AND COLOR BAR-CODING OF LARGE GENES |
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| Author(s): FLORIJN RJ, BONDEN LAJ, VROLIJK H, WIEGANT J, VAANDRAGER JW, BAAS F, DENDUNNEN JT, TANKE HJ, VANOMMEN GJB, RAAP AK |
| Source: HUMAN MOLECULAR GENETICS Volume: 4 Issue: 5 Pages: 831-836 Published: MAY 1995 |
| Times Cited: 94 References: 27 |
| Abstract: We have applied two-colour fluorescence in situ hybridization (FISH) to DNA fibers and combined it with digital imaging microscopy for the mapping of large cosmid contigs. The technique was validated using a set of unique plasmids and a cosmid contig both originating from the thyroglobulin (Tg) gene and previously mapped by restriction analysis. The resolution proved to be close to the theoretical lower limit of similar to 1 kb, ranging greater than or equal to 400 kb. Subsequently a 400 kb cosmid contig derived from a DMD-YAC was directly mapped by Fiber-FISH, The resulting map is in full agreement with the restriction map. Two-colour Fiber-FISH mapping thus showed to be capable for accurately sizing gaps and overlaps, and to identify chimeric or repeat sequence containing cosmids across a 400 kb region at once. The generated 400 kb 'colour bar-code' was subsequently used to map two DMD deletion breakpoints in patient DNA with an accuracy of 1-2 kb, The results underscore the value of this method for the delineation of chromosomal rearrangements for positional cloning and single patient clinical studies. |
| Document Type: Article |
| Language: English |
Addresses:
1. LEIDEN STATE UNIV, DEPT CYTOCHEM & CYTOMETRY, 2333 AL LEIDEN, NETHERLANDS
2. LEIDEN STATE UNIV, DEPT HUMAN GENET, 2333 AL LEIDEN, NETHERLANDS
3. UNIV AMSTERDAM, ACAD MED CTR, DEPT NEUROL, 1105 AZ AMSTERDAM, NETHERLANDS |
| Publisher: OXFORD UNIV PRESS UNITED KINGDOM, WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP |
| Subject Category: Biochemistry & Molecular Biology; Genetics & Heredity |
| IDS Number: QX780 |
| ISSN: 0964-6906 |
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