A rapid wire-based sampling method for DNA profiling.
This paper reports the results of a commission to develop a field deployable rapid short tandem repeat (STR)-based DNA profiling system to enable discrimination between tissues derived from a small number of individuals. Speed was achieved by truncation of sample preparation and field deployability by use of an Agilent 2100 Bioanalyser(TM). Human blood and tissues were stabbed with heated stainless steel wire and the resulting sample dehydrated with isopropanol prior to direct addition to a PCR. Choice of a polymerase tolerant of tissue residues and cycles of amplification appropriate for the amount of template expected yielded useful profiles with a custom-designed quintuplex primer set suitable for use with the Bioanalyser(TM). Samples stored on wires remained amplifiable for months, allowing their transportation unrefrigerated from remote locations to a laboratory for analysis using AmpFlSTR(®) Profiler Plus(®) without further processing. The field system meets the requirements for discrimination of samples from small sets and retains access to full STR profiling when required.
School of Biological Sciences, Flinders University,Adelaide, SA 5001, Australia.
Tong
T
Chen
David E A
DE
Catcheside
Alice
A
Stephenson
Chris
C
Hefford
K Paul
KP
Kirkbride
Leigh A
LA
Burgoyne
D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers; D019075:Electrophoresis, Capillary; D005838:Genotype; D006358:Hot Temperature; D006801:Humans; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D013048:Specimen Handling / Q000295:instrumentation* / Q000379:methods*; D013193:Stainless Steel
D017931:DNA Primers; D013193:Stainless Steel