Different whole-genome amplification methods as a preamplification tool in Y-chromosome Loci analysis.
Degraded and low template DNA is often analyzed in forensic genetics laboratories. Reliable analysis of degraded and low template DNA is of great importance, because its results impact the quality and reliability of expert testimonies. Recently, a number of whole-genome amplification (WGA) methods have been proposed as preamplification tools improving quantity and quality of DNA. We chose, investigated, and compared 7 WGA methods to evaluate their ability to "recover" degraded and nondegraded DNA. These methods include degenerate oligonucleotide primed polymerase chain reaction, primer extension preamplification (PEP) polymerase chain reaction, GenomePlex WGA (Sigma), multiple displacement amplification, GenomiPhi Amplification Kit (Amersham Biosciences), restriction and circularization aided rolling circle amplification, and blunt-end ligation-mediated WGA. Recently, we have described the comparison of these methods' efficiency and reliability using SGMPlus kit. However, Y-chromosome profiling is also often used in analysis of both nondegraded and degraded DNA. This includes criminal cases and investigation of kinship in male linage. Here we demonstrate the impact of WGA methods on Y-chromosome loci (Yfiler) reactivation.The best results for nondegraded DNA were obtained with GenomiPhi kit and PEP method. In the case of degraded DNA (200 base pairs), the most complete profiles were obtained with GenomePlex kit and PEP method. None of the analyzed methods allowed full reactivation of degraded (200 base pairs) DNA in terms of Y-chromosome loci profiling.
From the Medical University of Gdańsk, Institute of Forensic Medicine, Gdańsk, Poland.
Agnieszka
A
Maciejewska
Joanna
J
Jakubowska
Ryszard
R
Pawłowski
D041322:Chromosomes, Human, Y; D004247:DNA / Q000302:isolation & purification; D053836:DNA Degradation, Necrotic; D016172:DNA Fingerprinting / Q000379:methods; D056426:Genetic Loci; D015894:Genome, Human; D006801:Humans; D008297:Male; D018482:Muscle, Skeletal / Q000737:chemistry; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*
D004247:DNA