ABSTRACT
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.
Subject(s)
DNA Fingerprinting/methods , Electric Power Supplies , Equipment Failure , Polymerase Chain Reaction/methods , Electrophoresis, Agar Gel , Electrophoresis, Capillary , Humans , Specimen Handling , Tandem Repeat SequencesABSTRACT
The fingernails of Mary Sullivan, the last victim of the Boston Strangler, were examined to determine if any genetic information about the murderer could be obtained. The nails were extremely friable necessitating the development of new techniques for isolating and purifying DNA. DNA yields from nails were optimized by using a NaOH-based preparation technique, which was simple, efficient, and minimized handling. Methods for selectively and thoroughly removing exogenous material on nails were also developed through use of a species-specific PCR assay, wherein mitochondrial DNA from the nail could easily be differentiated from DNA of contaminating cells.