Abstract
Purpose : To apply capillary electrophoresis for rapid screening for Y microdeletions.
Methods : A set of 25 specific sequence tagged sites that cover the azoospermia factor a, b, and c regions of the Y chromosome was amplified in 5 fluorescent multiplex sets each including 5 primer pairs. One of the primers of each pair was labeled with a fluorescent tag attached to the 5′-end. After PCR amplification, analysis of the obtained PCR products was performed using capillary electrophoresis (ABI Prism 3100 Genetic Analyzer). The method was employed to determine Y microdeletions in azoospermic (n = 49) and severe oligozoospermic (n = 149) men.
Results : The number of PCR cycle (from 45 to 30) and the amount of DNA template (20-fold) used in fluorescent multiplex PCR were reduced because of the high sensitivity of capillary electrophoresis. Approximately 1000 multiplex PCR sets from 198 patients were analyzed simultaneously within 50 h. Y microdeletions were found in 3 out of the 198 azoospermic or severe oligozoospermic men.
Conclusions : Application of capillary electrophoresis for detection of PCR products provides a semiautomated, high throughput method for rapid screening for microdeletions on the Y chromosome.
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Bor, P., Hindkjær, J., Kølvraa, S. et al. A New Approach for Screening for Y Microdeletions: Capillary Electrophoresis Combined with Fluorescent Multiplex PCR. J Assist Reprod Genet 20, 46–51 (2003). https://doi.org/10.1023/A:1021215006775
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DOI: https://doi.org/10.1023/A:1021215006775