PCR-amplification and detection of the human D1S80 VNTR locus. Amplification conditions, population genetics and application in forensic analysis.
A series of experiments has been performed to evaluate amplification and typing of the D1S80 VNTR locus. The validation study that has been carried out showed that correct D1S80 typing results can be obtained when a defined amplification protocol and a high-resolution polyacrylamide gel electrophoresis method are used. The use of the Chelex extraction protocol has substantially reduced the processing time. DNA-extraction, amplification and subsequent typing can be performed in one day. The discrimination power of this locus is 0.94 in a Dutch Caucasian population sample. The system is extremely sensitive: 0.1 ng of genomic DNA gave a correct typing result. The test could also detect the correct genotypes in mixed samples containing DNA from different individuals. Even if the major type was in a 20-fold excess, the minority type could still be amplified and typed correctly. We have found no deviation from Hardy-Weinberg equilibrium in a Dutch Caucasian population sample. Evidence for the somatic stability of this locus was obtained from a set of experiments where we compared DNA-profiles from corresponding blood, semen and saliva samples. The results of this study suggest that in the near future analysis of the D1S80 locus by DNA-amplification can be applied in actual forensic case work.
Kloosterman
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Gerechtelijk Laboratorium van het Ministerie van Justitie, Rijswijk, The Netherlands.
Budowle
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Daselaar
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D000483:Alleles; D002874:Chromosome Mapping; D003431:Cross-Cultural Comparison; D004247:DNA / Q000235:genetics*; D005260:Female; D005554:Forensic Medicine; D005784:Gene Amplification / Q000235:genetics*; D005787:Gene Frequency / Q000235:genetics; D006580:Genetic Carrier Screening; D005819:Genetic Markers / Q000235:genetics; D005828:Genetics, Population; D006801:Humans; D008297:Male; D009426:Netherlands; D016133:Polymerase Chain Reaction / Q000379:methods*; D011180:Postmortem Changes; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*
D005819:Genetic Markers; D004247:DNA