An Alu-based, MGB Eclipse real-time PCR method for quantitation of human DNA in forensic samples.
The forensic community needs quick, reliable methods to quantitate human DNA in crime scene samples to replace the laborious and imprecise slot blot method. A real-time PCR based method has the possibility of allowing development of a faster and more quantitative assay. Alu sequences are primate-specific and are found in many copies in the human genome, making these sequences an excellent target or marker for human DNA. This paper describes the development of a real-time Alu sequence-based assay using MGB Eclipse primers and probes. The advantages of this assay are simplicity, speed, less hands-on-time and automated quantitation, as well as a large dynamic range (128 ng/microL to 0.5 pg/microL).
Nicklas
Janice A
JA
Vermont Forensic Laboratory, Department of Public Safety, 103 S. Main St., Waterbury, VT 05671, USA.
Buel
Eric
E
D020087:Alu Elements; D000818:Animals; D052160:Benzothiazoles; D004247:DNA / Q000032:analysis*; D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers; D015342:DNA Probes; D003959:Diamines; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D008297:Male; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction / Q000379:methods*; D011804:Quinolines; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D020080:Tandem Repeat Sequences
D052160:Benzothiazoles; D017931:DNA Primers; D015342:DNA Probes; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D004247:DNA