Summary
The polymorphism of the D1S80 locus has been analyzed in a population sample of 203 unrelated individuals living in Madrid (central Spain) by PCR and subsequent semi-dry discontinuous polyacrylamide gel electrophoresis (Tris-chloride/Tris-glycine buffer system) followed by silver staining, The electrophoretic system described in this study offers high resolution in the separation of the different D1S80 alleles allowing the detection of microvariability around the allele T22 in the spanish population. Twenty different alleles containing 17–40 repeats of the basic 16bp unit were distinguished. The alleles T18 and T24 were found to be relatively common in Spain, as in other populations:, with frequencies of 0.224 and 0.372, respectively. No evidence of significant deviations from Hardy-Weinberg equilibrium was found in these preliminary population data.
Zusammenfassung
Der Polymorphismus des Locus D1S80 wurde in einer Bevölkerungsstichprobe von 203 unverwandten, in Madrid lebenden Personen unter-sucht. Die Untersuchung erfolgte mit Hilfe der PCR und nachfolgender, halbtrockener diskontinuierlicher Polyacrylamidgel-Elektrophorese (Tris-chlorid/Tris-glycin-Puffersystem) mit nachfolgender Silberfärbung. Das beschriebene elektrophoretische System dieser Untersuchung bietet eine hohe Auflösung in der Auftrennung der verschiedenen D1S80-Allele und erlaubt den Nachweis einer Mikrovariabilität um das Allel T22 in der spanischen Bevölkerung, welche zwischen 17 und 40 Wiederholungseinheiten der zugrunde liegenden 16 Bp-Einheit enthalten. Die Allele T18 and T24 wurden als relativ häufig in Spanien gefunden, wie auch in anderen Populationen, mit Frequenzen von 0,224 bzw. 0,372. Es fand sich kein Beweis für signifikante Abweichungen vom Hardy-Weinberg-Gleichgewicht in dieser vorläufigen Populationsstudie.
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References
Allen, RC, Graves G, Budowle G (1989) Polymerase chain reaction amplification products separated on rehydratable polyacrylamide gels and stained with silver. Bio Techniques 7:736–744
Blum H, Beier H, Gross HJ (1987) Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8:93–99
Boerwinkle E, Xiong W, Fourest E, Chan L (1989) Rapid typing of tandemly repeated hypervariable loci by the polymerase chain reaction: application to the apolipoprotein B 3′hypervariable region. Proc Natl Acad Sci USA 86:212–216
Brenner C, Morris JW (1990) Paternity index calculations in single locus hypervariable DNA probes: validation and other studies. In: Preceedings for the International Symposium of Human Identification, Promega corporation, Madison, USA
Budowle B, Chakraborty R, Giusti AM, Eisenberg AJ, Allen RC (1991) Analysis of the variable number of tandem repeats locus D1S80 by the polymerase chain reaction followed by high resolution polyacrylamide gel electrophoresis. Am J Hum Genet 48:137–144
Budowle B, Baechtel FS, Comey CT (1992) Some considerations for use of AMP-FLPs for identity testing. In: Rittner C, Schneider PM (eds) Advances in forensic haemogenetics 4. Springer, Berlin Heidelberg New York, pp 12–17
Caskey CT; Hammond HA (1992) Forensic use of short tandem repeats via PCR. In: Rittner C, Schneider PM (eds) Advances in forensic haemogenetics 4. Springer, Berlin Heidelberg New York, pp 18–25
Davis BJ (1964) Disc electrophoresis. II. Method and application to human serum proteins. Ann NY Acad Sci 121:404–427
Horn GT, Richards B, Klinger KW (1989) Amplification of a highly polymorphic VNTR segment by the polymerase chain reaction. Nucleic Acids Res 17:21–40
Kasai K, Nakamura Y, White RL (1990) Amplification of a variable number of tandem repeats (VNTR) locus (pMCT118) by the polymerase chain reaction (PCR) and its application to forensic science. J Forensic Sci 35:1196–1200
Ludwig EH, Friedl W, McCarthy BJ (1989) High-resolution analysis of a hypervariable region in the human apolipoprotein B gene. Am J Hum Genet 45:458–464
Olsson I, Stalberg R (1986) A new semi-dry method for horizontal electrophoresis in ultrathin polyacrylamide gels. In: Dunn MJ (ed) Electrophoresis'86. VCH, Weinheim, pp 230–232
Ornstein L (1964) Disc electrophoresis. I. Background and theory. Ann NY Acad Sci 121:321–349
Priestley L, Kumar D, Sykes B (1990) Amplification of the COL2A1 3′ variable region used for segregation analysis in a family with Stickler syndrome. Hum Genet 85:525–526
Rand S, Puers C, Skowasch K, Wiegand P, Budowle B, Brinkmann B (1992) Population genetics and forensic efficiency data of 4 AMPFLP's. Int J Leg Med 104:329–333
Sajantila A, Budowle B, Ström M, Johnsson V, Lukka M, Peltonen L, Ehnholm C (1992) PCR amplification of alleles at theD1S80 locus: comparison of a Finnish and a North American caucasian population sample, and forensic casework evaluation. Am J Hum Genet 50: 816–825
Smith JC, Anwar R, Riley J, Jenner D, Markham AF, Jeffreys AJ (1989) Highly polymorphic minisatelite sequences: allele frequencies and mutation rates for five locus-specific probes in a Caucasian population. J Forensic Sci Soc 30:19–32
Wolff RK, Nakamura YR, White R (1988) Molecular characterization of a spontaneously generated new allele at a VNTR locus: no exchange of flanking DNA sequence. Genomics 3:347–351
Wu S, Seino S, Bell GI (1990) Human collagen, type II, alpha 21, (COL2A1) gene: VNTR polymorphism detected by gene amplification. Nucleic Acids Res 18:3102
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Alonso, A., Martin, P., Albarran, C. et al. Amplified fragment length polymorphism analysis of the VNTR locus D1S80 in central Spain. Int J Leg Med 105, 311–314 (1993). https://doi.org/10.1007/BF01222113
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DOI: https://doi.org/10.1007/BF01222113