title: Evaluation of a co-extraction method for real-time PCR-based body fluid identification and DNA typing.
abstract: Body fluid identification and individual identification are an important series of tests in usual criminal investigations. Recent reports have demonstrated a new approach using DNA/RNA co-extraction methods in which RNA for body fluid identification and DNA for short tandem repeat (STR) typing are extracted simultaneously from the same sample. This study evaluated a standard co-extraction kit, the AllPrep® DNA/RNA Mini Kit, in order to demonstrate the availability of the co-extraction procedure for those real-time polymerase chain reaction-based body fluid identification methods that we have validated previously. We demonstrated that the use of the Allprep Kit, for which we adjusted the lysis temperature to 56°C to improve extraction efficiency, can simultaneously extract sufficient RNA and DNA for body fluid identification and STR analysis; however, a longer incubation at a high temperature slightly affected the ΔCt value of each target gene and appeared to be not as effective for DNA extraction from old stains as from 1-day-old stains. This method is promising for future forensic investigations because the use of this kit can reduce sample consumption for body fluid identification and DNA typing.
authors.affiliation: National Research Institute of Police Science, Chiba 277-0882, Japan. Electronic address: k-watanabe@nrips.go.jp.
authors.forename: Ken
authors.initials: K
authors.lastname: Watanabe
authors.affiliation: National Research Institute of Police Science, Chiba 277-0882, Japan.
authors.forename: Yasuki
authors.initials: Y
authors.lastname: Iwashima
authors.affiliation: National Research Institute of Police Science, Chiba 277-0882, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Akutsu
authors.affiliation: National Research Institute of Police Science, Chiba 277-0882, Japan.
authors.forename: Kazumasa
authors.initials: K
authors.lastname: Sekiguchi
authors.affiliation: National Research Institute of Police Science, Chiba 277-0882, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Sakurada
mesh_terms: D001826:Body Fluids; D016172:DNA Fingerprinting / Q000379:methods*; D006801:Humans; D018895:Microsatellite Repeats; D060888:Real-Time Polymerase Chain Reaction
keywords: Body fluid identification; DNA/RNA co-extraction; Real-time RT-PCR