The successful DNA typing of samples following a thermal cycler power loss.

Autor: Kokoszka JE; Orchid Cellmark, 20271 Goldenrod Lane, Germantown, MD 20876, USA., Cline RE, Leisy C, Grossweiler LL, Word CJ
Jazyk: angličtina
Zdroj: Journal of forensic sciences [J Forensic Sci] 2006 Sep; Vol. 51 (5), pp. 1074-9.
DOI: 10.1111/j.1556-4029.2006.00240.x
Abstrakt: An approach for generating DNA profiles when critical samples have been consumed and a power outage occurs during the polymerase chain reaction (PCR) amplification reaction is described. This study demonstrates that a complete and accurate DNA short tandem repeat profile can be obtained: (1) when single source DNA samples are amplified for 26, 27, or 28 cycles using the Profiler Plus and COfiler Amplification Kits after an interruption in amplification, (2) from mock samples when PCR amplification has been interrupted early (after five cycles) or late (after 18 cycles) and the sample is subjected to an additional round of amplification, even after incubation of the sample at room temperature overnight, and (3) from nonprobative casework samples interrupted after approximately 18 cycles of amplification, an overnight incubation at room temperature and subjected to one or two additional rounds of PCR amplification for approximately 26 total cycles. Samples interrupted before five completed cycles and subjected to additional PCR cycles yielded variable results.
Databáze: MEDLINE