title: DNA probe pooling for rapid delineation of chromosomal breakpoints.
abstract: Structural chromosome aberrations are hallmarks of many human genetic diseases. The precise mapping of translocation breakpoints in tumors is important for identification of genes with altered levels of expression, prediction of tumor progression, therapy response, or length of disease-free survival, as well as the preparation of probes for detection of tumor cells in peripheral blood. Similarly, in vitro fertilization (IVF) and preimplantation genetic diagnosis (PGD) for carriers of balanced, reciprocal translocations benefit from accurate breakpoint maps in the preparation of patient-specific DNA probes followed by a selection of normal or balanced oocytes or embryos. We expedited the process of breakpoint mapping and preparation of case-specific probes by utilizing physically mapped bacterial artificial chromosome clones. Historically, breakpoint mapping is based on the definition of the smallest interval between proximal and distal probes. Thus, many of the DNA probes prepared for multiclone and multicolor mapping experiments do not generate additional information. Our pooling protocol, described here with examples from thyroid cancer research and PGD, accelerates the delineation of translocation breakpoints without sacrificing resolution. The turnaround time from clone selection to mapping results using tumor or IVF patient samples can be as short as 3 to 4 days.
authors.affiliation: Department of Chemical and Materials Engineering, National Chin-Yi University of Technology, Taiping City, Taichung, Taiwan, Republic of China.
authors.forename: Chun-Mei
authors.initials: CM
authors.lastname: Lu
authors.forename: Johnson
authors.initials: J
authors.lastname: Kwan
authors.forename: Adolf
authors.initials: A
authors.lastname: Baumgartner
authors.forename: Jingly F
authors.initials: JF
authors.lastname: Weier
authors.forename: Mei
authors.initials: M
authors.lastname: Wang
authors.forename: Tomas
authors.initials: T
authors.lastname: Escudero
authors.forename: Santiago
authors.initials: S
authors.lastname: Munné
authors.forename: Horst F
authors.initials: HF
authors.lastname: Zitzelsberger
authors.forename: Heinz-Ulrich G
authors.initials: HU
authors.lastname: Weier
mesh_terms: D002460:Cell Line; D019457:Chromosome Breakage; D022202:Chromosomes, Artificial, Bacterial; D002878:Chromosomes, Human, Pair 1; D002882:Chromosomes, Human, Pair 13; D002894:Chromosomes, Human, Pair 4; D003001:Cloning, Molecular; D020451:Contig Mapping; D015342:DNA Probes; D005260:Female; D006801:Humans; D008297:Male; D008677:Metaphase; D011247:Pregnancy; D019836:Preimplantation Diagnosis; D013964:Thyroid Neoplasms / Q000235:genetics; D014178:Translocation, Genetic; D055815:Young Adult
chemical_list: D015342:DNA Probes