(Received 12 March 2005; accepted 23 April 2005)
Published Online: August
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EDTA blood samples were collected from 200 unrelated Tunisian healthy adults coming from different regions of the country. All subjects gave their informed consent. DNA was extracted from leucocytes by the standard phenol chloroform technique. Ampli.- cation of the D1S80 was carried out by polymerase chain reaction (PCR) using the primers described by Kasai et al. (1). Reaction was performed in a total volume of 50 µL containing 200 ng genomic DNA, 10 mM Tris-Hcl (pH 8.3), 50 mM Kcl, 1.5 mM MgC12, 2.5 units of AmplitaqTM polymerase (Perkin Elmer-Cetus USA), 200 µMdNTP and 1µM of each primer. The thermocycling used GeneAmpR PCR System 9700 PE Applied Biosystems. PCR fragments were analysed using a 3% agarose gel electrophoresis (Agarose LE Promega, France); a D1S80 allelic ladder (Perkin Elmer Cetus) was applied on the gel at two-lane intervals to allow correct sizing of the ampli.ed fragments. DNA fragments were visualised by ethidium bromide staining. Statistical analysis was performed using The TFPGA program version 1.3 (2).
Faculté de Médecine de Tunis,
Service Maladies Héréditaires Hôpital Charles Nicolle, Tunis,
Stock #: JFS2005185