Premium
Hybridization and polymerase chain reaction amplification of simple repeated DNA sequences for the analysis of forensic stains
Author(s) -
Roewer Lutz,
Rieß Olaf,
Prokop Otto
Publication year - 1991
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150120214
Subject(s) - polymerase chain reaction , hypervariable region , oligonucleotide , multiple displacement amplification , dna , biology , microbiology and biotechnology , stain , dna nanoball sequencing , genomic dna , dna sequencing , genetics , computational biology , gene , dna extraction , genomic library , staining , base sequence
We have evaluated oligonucleotide hybridization and amplification techniques with regard to quantity and quality of genomic DNA that is under investigation in practical forensic case work. In order to obtain sufficient information from analyzing stain material, we use hypervariable simple repeat sequences for individualization, which occur in all eukaryotic genomes. For the analysis of larger amounts of stains (> 500 ng DNA) the multilocus probes (CAC) 5 /(GTG) 5 Probes are written in capitals, genomic noncoding sequences in lower case are superior because of their discrimination potential – provided that the hybridizing DNA is of high molecular weight. The less discriminating probes (CT) 8 and (GACA) 4 are more sensitive (minimal amount: 100ng DNA) and still informative when the DNA is degraded. To increase the sensitivity of forensic stain analysis in special cases we have used the polymerase chain reaction technique to amplify hypervariable simple (gt) n /(ga) m repeat structures from the intron 2 of HLA ‐ DRB genes. Largely independent of the starting amount of DNA and independent of the degradation status, we were able to generate discriminating DNA fragments, which can be used to type (i) microstains and (ii) totally degraded material including human mummy DNA.