Assay model, design of MLPA telomeric and centromeric probe sets, and verification of results obtained using MLPA.

<p>A. Overview of the chromosome V +Ty912 assay strain. The <i>Ty912</i> was inserted between <i>NPR2</i> and <i>CIN8</i> oriented so that it was transcribed towards the telomere. <i>Ty912</i> and other genes are not drawn to scale. B. Schematic of the MLPA telomeric and centromeric probes. Telomeric and centromeric probes are designated by solid black and grey lines above the chromosomes, respectively. Ty delta elements are represented as hollow blue triangles, and Ty1 and Ty2 elements are designated as pairs of hollow blue triangles connected by solid blue lines. The orientation of the triangles represent the transcriptional orientation of the elements. Tys and chromosome arms are not drawn to scale. C. Schematic of the MLPA process (1: chromosomal DNA, 2: MLPA probes, 3: universal PCR primers). MLPA probes (2) are hybridized to the chromosomal DNA (1), ligated, amplified by PCR using universal primers (3) and analyzed using capillary electrophoresis. D. Graphical display of MLPA data generated using telomeric MLPA probes. Chromosome arms are represented by peaks in either a wild-type isolate or GCR-containing isolate. Ratios of the normalized peak areas compared to a relative set of wild-type control isolates are given above the indicated peaks of interest. Black vertical arrows indicate either a chromosome V-L deletion (left arrow) or a chromosome XIII-L duplication (right arrow) in the GCR-containing isolate and normal chromosome arm complements in the wild-type isolate.