A rapid typing system at three STR loci from bloodstains using a simple DNA extraction kit and capillary electrophoresis.
A rapid typing method for STR analysis from bloodstains was devised. DNA from three-year-old bloodstains of six individuals was extracted with a cationic detergent in a newly-released kit. Amounts of DNA extracted by the kit were 16.2 +/- 1.8 ng from 10 microliters of fresh blood samples. DNA was also extracted from bloodstains of three months and three years left at room temperature, and amounts of extracted DNA were estimated at 2.2 +/- 1.0 ng and 0.1 +/- 0.1 ng, respectively. The PCR mixture supplied with the kit, plus three sets of fluorescently labeled primers, were directly added to the tube used for DNA extraction. The amplified products were then analyzed by capillary electrophoresis, and the obtained data were automatically sized and typed using computer software. Three genotypes of all bloodstains except one sample at a single locus were determined correctly as those of freshly collected blood from the same individuals although some stains contained no more than 0.1 ng. Using this typing system, we were able to type three STR loci, HUMTH01, HUMF13A1 and HUMFES/FPS, from a single bloodstain within five hours. This typing is particularly useful for forensic practice.
Yamamoto
T
T
Department of Legal Medicine, Nagoya University School of Medicine.
Kojima
T
T
Nozawa
H
H
Huang
X L
XL
Ohtaki
H
H
Uchihi
R
R
Tamaki
K
K
Katsumata
Y
Y
D001801:Blood Stains; D004247:DNA / Q000302:isolation & purification*; D019075:Electrophoresis, Capillary; D006801:Humans; D016133:Polymerase Chain Reaction; D011933:Reagent Kits, Diagnostic / Q000592:standards*; D012091:Repetitive Sequences, Nucleic Acid
D011933:Reagent Kits, Diagnostic; D004247:DNA