Cryptic Translocation Identification in Human and Mouse using Several Telomeric Multiplex FISH (TM-FISH) Strategies

Henegariu, O ; Artan, S ; Greally, J M ; Chen, X-N ; Korenberg, J R ; Vance, G H ; Stubbs, L ; Bray-Ward, P ; Ward, D C

Lawrence Livermore National Laboratory

关键词: Chromosomes;    Personnel;    Fluorescence;    Human Chromosomes;    Adobe;   

DOI  :  10.2172/15004910
RP-ID  :  UCRL-ID-155039
RP-ID  :  W-7405-ENG-48
RP-ID  :  15004910

美国|英语

来源: UNT Digital Library

【 摘 要 】

Experimental data published in recent years showed that up to 10% of all cases with mild to severe idiopathic mental retardation may result from small rearrangements of the subtelomeric regions of human chromosomes. To detect such cryptic translocations, we developed a ''telomeric'' multiplex FISH assay, using a set of previously published and commercially available subtelomeric probes. This set of probes includes 41 cosmid/PAC/P1 clones located from less than 100kb to about 1 Mb from the end of the chromosomes. Similarly, a published mouse probe set, comprised of BACs hybridizing to the closest known marker toward the centromere and telomere of each mouse chromosome, was used to develop a mouse-specific ''telomeric'' M-FISH. Three different combinatorial labeling strategies were used to simultaneously detect all human sub-telomeric regions on one slide. The simplest approach uses only three fluors, and can be performed in laboratories lacking sophisticated imaging equipment or personnel highly trained in cytogenetics. A standard fluorescence microscope equipped with only three filters is sufficient. Fluor-dUTPs and labeled probes can be custom-made, thus dramatically reducing costs. Images can be prepared using generic imaging software (Adobe Photoshop), and analysis performed by simple visual inspection.