Balkan Journal of Medical Genetics

PRELIMINARY FINDINGS OF α-ACTININ-3 GENE DISTRIBUTION IN ELITE TURKISH WIND SURFERS
Ulucan K1, Göle S1, Altindas N2, Güney AI3
*Corresponding Author: Dr. Korkut Ulucan, Department of Molecular Biology and Genetics, Faculty of Engineering and Natural Sciences, Üsküdar University, Altunizade Mahallesi, Haluk Turksoy Sok., No. 14 Uskudar, Istanbul 34662, Turkey; +90-216-400-2222-2409, +90-216-474-1256; korkutulucan@hotmail.com
page: 69

MATERIALS AND METHODS

Eight National Team wind surfers and medal winners, were enrolled in an ACTN3 genotyping procedure. The study procedure was in accordance with the principles of the Declaration of Helsinki II and all subjects provided written informed consent prior to enrollment. DNA isolation was carried out by using high pure polymerase chain reaction (PCR) Template Preparation Kit (Roche Diagnostics, Mannheim, Germany). The region of interest of the gene was amplified by using the following primers: forward, 5’-CTG TTG CCT GTG GTA AGT GGG-3’ and reverse, 5’-TGG TCA CAG TAT GCA GGA GGG-3’. Polymerase chain reaction was performed by initial denaturation at 95°C for 5 min., followed by 35 cycles of denaturation at 95°C for 30 seconds, annealing and extension at 72°C for 1 min., and a final extension for 7 min. at 72°C. The 290 bp length amplicons were digested by DdeI (New England Biolabs, Beverly, MA, USA) in a condition recommended by manufacturer. The wild-type allele, 577R, showed fragments of 205 and 85 bp on 10.0% polyacrylamide gel electrophoresis (PAGE), whereas the variant allele, 577X, showed fragments of 108, 97 and 85 bp. Visualization of the digested fragments was performed by staining with ethidium bromide.

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