Application of FTA technology to extraction of sperm DNA from mixed body fluids containing semen.
FTA technology is a novel method designed to simplify the collection, shipment, archiving and purification of nucleic acids from a wide variety of biological sources. In this study, we report a rapid and simple method of extracting DNA from sperm when body fluids mixed with semen were collected using FTA cards. After proteinase K digestion of the sperm and body fluid mixture, the washed pellet suspension as the sperm fraction and the concentrated supernatant as the epithelial cell fraction were respectively applied to FTA cards containing DTT. The FTA cards were dried, then directly added to a polymerase chain reaction (PCR) mix and processed by PCR. The time required from separation of the mixed fluid into sperm and epithelial origin DNA extractions was only about 2.5-3h. Furthermore, the procedure was extremely simple. It is considered that our designed DNA extraction procedure using an FTA card is available for application to routine work.
Fujita
Yoshihiko
Y
Department of Forensic Medicine, Institute of Health Biosciences, The University of Tokushima Graduate School, 3-8-15 kuramoto, Tokushima 770-8503, Japan.
Kubo
Shin-ichi
S
D004247:DNA / Q000302:isolation & purification*; D016172:DNA Fingerprinting; D019286:Endopeptidase K; D005260:Female; D005838:Genotype; D006801:Humans; D008297:Male; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods*; D016133:Polymerase Chain Reaction; D011902:Rape; D012463:Saliva / Q000378:metabolism; D013094:Spermatozoa / Q000378:metabolism*; D020080:Tandem Repeat Sequences; D014621:Vagina / Q000378:metabolism; D014867:Water
D014867:Water; D004247:DNA; D019286:Endopeptidase K