Rational design of landmark probes for quantitative DNA fiber mapping (QDFM)

AUTOR(ES)

Hsieh, Huangpin B.

FONTE

Oxford University Press

RESUMO

Rapid construction of high-resolution physical maps requires accurate information about overlap between DNA clones and the size of gaps between clones or clone contigs. We recently developed a procedure termed ‘quantitative DNA fiber mapping’ (QDFM) to help construct physical maps by measuring the overlap between clones or the physical distance between non-overlapping contigs. QDFM is based on hybridization of non-isotopically labeled probes onto DNA molecules that were bound to a solid support and stretched homogeneously to ~2.3 kb/µm. In this paper, we describe the design of probes that bind specifically to the cloning vector of DNA recombinants to facilitate physical mapping. Probes described here delineate the most frequently used cloning vectors such as BACs, P1s, PACs and YACs. As demonstrated in representative hybridizations, vector-specific probes provide valuable information about molecule integrity, insert size and orientation as well as localization of hybridization domains relative to specifically-marked vector sequences.

Documentos Relacionados

• Rational Design of DNA Sequence-Based Strategies for Subtyping Listeria monocytogenes
• Rational design of DNA sequences for nanotechnology, microarrays and molecular computers using Eulerian graphs
• The Design of Rational Combination Chemotherapy for Cancer
• Rational design of diagnostic and vaccination strategies for tuberculosis
• Sequence-Based Design of Single-Copy Genomic DNA Probes for Fluorescence In Situ Hybridization