Construction of two near-kilobase resolution restriction maps of the 5' regulatory region of the human apolipoprotein B gene by quantitative DNA fiber mapping (QDFM)

Cytogenetics and Cell Genetics
T DuellH U Weier

Abstract

Quantitative DNA fiber mapping (QDFM) is a high-resolution technique for physical mapping of DNA. The method is based on hybridization of fluorescently labeled DNA probes to individual DNA molecules stretched on a chemically modified glass surface. We now demonstrate and validate a rapid QDFM-based approach for the mapping of multiple restriction sites and precise localization of restriction fragments in large genomic clones. Restriction fragments of a 70-kb P1 clone (P1-70) containing the 5' region of the human apolipo-protein B gene (APOB) were subcloned and mapped along straightened P1-70 DNA molecules. Multicolor fluorescence in situ hybridization (FISH) and digital image analysis allowed us to rapidly position 29 restriction fragments, ranging in size from 0.5 kb to 8 kb, and to map 43 restriction sites. The restriction map obtained by QDFM was in excellent agreement with information obtained by RecA-assisted restriction endonuclease (RARE) cleavage, long-range PCR, and DNA sequence analyses of the P1-70 clone. These data demonstrate that QDFM is a rapid, reliable method for detailed restriction site-mapping of large DNA clones.

Citations

Mar 29, 2000·Nucleic Acids Research·H B HsiehH U Weier
Dec 26, 2008·Molecular Cytogenetics·Karin M Greulich-BodeHeinz-Ulli G Weier
Dec 10, 2009·Folia Histochemica Et Cytobiologica·Johnson KwanHeinz-Ulrich G Weier
Jul 18, 2001·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·H U Weier
Feb 2, 2013·Chemical Communications : Chem Comm·Jinyong LeeKyubong Jo

❮ Previous
Next ❯

Related Concepts

Related Feeds

Artificial Chromosomes

Artificial chromosomes are genetically engineered chromosomes derived from the DNA of a species. Discover the latest research on artificial chromosomes here.