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    Volume 48, Issue 2 (March 2003)

    Variation of the Hypervariable Region-1 of Mitochondrial DNA in Central-Eastern Italy

    Published Online: March


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    DNA samples were extracted from fresh blood (5 mL in EDTA tube) by alkali bursting of blood cells followed by protein digestion with proteinase K. After a phenol-chloroform extraction, DNA was precipitated with absolute ethanol and dried at room temperature. The DNA pellet was diluted in 100 μL of TBE buffer (Tris HCl, 10mM, pH 7.5; EDTA 1mM). PCR amplification was carried out by using a Perkin Elmer 2400 thermal cycler in 25 μL reaction volume. The product of amplification was purified with a High Pure Product Purification Kit (Roche Diagnostics GmbH, Mannheim, Germany). The sequencing reactions were performed separately on each strand with the primers L15990 and H16401. The hypervariable region-1 (HVR1) sequence analysis was carried out using the A.L.F. semi-automated sequencer (Pharmacia Biotech, Uppsala, Sweden). We observed a total of 44 different haplotypes out of 50 individuals examined, 41 of which were singletons. Only 8 haplotypes (no. 09, 13, 29, 39, 40, 42, 44, and 50) have been already observed among the Italian populations database available at The parameters of intrapopulation diversity (haplotype diversity 0.990 ± 0.008; mean number of pairwise differences 4.979 ± 2.463) fall into the range of values reported for Italian populations (1). The values of Tajima (1,944, p = 0.015) and Fu's tests (-25.552, p = 0.00) are those expected for populations who have had a large and sudden expansion (2). Applying an exact test for population comparisons implemented in the Arlequin software (3), we obtained statistically significant p-values for the comparison between our sample and those from Tuscany (0.004) Sardinia (0.039). Complete data are available at the e-mail address of the corresponding author upon request.

    Stock #: JFS2002350

    ISSN: 0022-1198

    DOI: 10.1520/JFS2002350

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    Title Variation of the Hypervariable Region-1 of Mitochondrial DNA in Central-Eastern Italy
    Symposium , 0000-00-00
    Committee E30