STR DNA typing: increased sensitivity and efficient sample consumption using reduced PCR reaction volumes.
Improvements in detection limits/sensitivity and lower sample consumption are potential benefits of reducing PCR reaction volumes used in forensic DNA typing of crime scene samples. This premise was studied first with experimental mixtures and a nine-loci megaplex, which demonstrated stochiometric amplification and accurate detection. Next, adjudicated casework samples were subjected to amplification under 15 different template DNA to PCR reaction volume ratios. Reduction of PCR reaction volume and DNA down to 10 microL and 0.500 ng, respectively, produced identical profiles with the same signal intensity and heterozygous allele peak height ratio (HR). Reduction to 5 microL and 0.063 ng yielded HR values that were slightly affected in one to three STR loci. PCR reaction volume reduction can enhance detection and sensitivity while reducing the consumption of irreplaceable crime scene samples.
Leclair
Benoît
B
National DNA Data Bank, Royal Canadian Mounted Police, 1200 Vanier Parkway, Ottawa, Ontario, Canada.
Sgueglia
Joanne B
JB
Wojtowicz
Patricia C
PC
Juston
Ann C
AC
Frégeau
Chantal J
CJ
Fourney
Ron M
RM
D016022:Case-Control Studies; D004247:DNA / Q000032:analysis*; D016172:DNA Fingerprinting / Q000379:methods*; D004586:Electrophoresis; D005240:Feasibility Studies; D005260:Female; D005554:Forensic Medicine / Q000379:methods; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D020080:Tandem Repeat Sequences
D004247:DNA