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Optimized DNA extraction to improve reproducibility of short tandem repeat genotyping with highly degraded DNA as target
Author(s) -
Schmerer Wera M.,
Hummel Susanne,
Herrmann Bernd
Publication year - 1999
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/(sici)1522-2683(19990101)20:8<1712::aid-elps1712>3.0.co;2-6
Subject(s) - genotyping , reproducibility , dna extraction , microsatellite , dna , polymerase chain reaction , str analysis , extraction (chemistry) , biology , chromatography , chemistry , genotype , genetics , gene , allele
The reproducibility of short tandem repeat (STR) genotyping of highly degraded DNA is often reduced due to artifacts generated during polymerase chain reaction (PCR) amplification. The frequency and amount of these artifacts are related to the quality and quantity of the DNA amplified. Consequently, the aim of this investigation was the optimization of DNA extraction to increase the reproducibility of STR genotyping of samples containing highly degraded DNA. Starting from a standard extraction protocol, systematic variation of individual parameters resulted in optimized protocols for three categories of ancient human bone material (different degrees of DNA degradation) and a consensus protocol for the extraction of a broad range of ancient DNA preservation states.