Preparation of degraded human DNA under controlled conditions.
DNA typing through analysis of short tandem repeats (STRs) and mitochondrial DNA (mtDNA) by means of the polymerase chain reaction (PCR) and sequencing are the common methods for the forensic identification of persons and reconstruction of kinship, especially when skeletal human remains have to be analyzed. Furthermore, samples typically found at crime scenes may be both quantitatively and qualitatively inadequate since they may contain very scarce and often degraded DNA due to exposure to heat, light, humidity, and microorganisms. In order to improve the performance of STR typing technology in those cases where DNA availability is limited, it would be desirable to have a source of degraded DNA with known properties. For this purpose, we have developed a method to prepare artificially degraded DNA under controlled conditions. By treatment of genomic DNA with sonication and DNAse I we have produced DNA fragments within a defined range of lengths. STR typing of this degraded DNA with a commercially available multiplex kit could only produce partial profiles as indicated by the absence of STR alleles with sizes >200 bp. This artificially degraded DNA can be used for the improvement and standardization of STR typing protocols when only highly degraded DNA is available for analysis.
Bender
Klaus
K
Institute of Legal Medicine, Johannes Gutenberg University, Am Pulverturm 3, D-55131 Mainz, Germany. kbender@mail.uni-mainz.de
Farfán
María José
MJ
Schneider
Peter M
PM
D016172:DNA Fingerprinting / Q000592:standards*; D053938:DNA Fragmentation; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000235:genetics*; D003850:Deoxyribonuclease I; D005260:Female; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction; D013010:Sonication; D020080:Tandem Repeat Sequences
D017931:DNA Primers; D004272:DNA, Mitochondrial; D003850:Deoxyribonuclease I