Establishing a novel automated magnetic bead-based method for the extraction of DNA from a variety of forensic samples.
Automated systems have been increasingly utilized for DNA extraction by many forensic laboratories to handle growing numbers of forensic casework samples while minimizing the risk of human errors and assuring high reproducibility. The step towards automation however is not easy: The automated extraction method has to be very versatile to reliably prepare high yields of pure genomic DNA from a broad variety of sample types on different carrier materials. To prevent possible cross-contamination of samples or the loss of DNA, the components of the kit have to be designed in a way that allows for the automated handling of the samples with no manual intervention necessary. DNA extraction using paramagnetic particles coated with a DNA-binding surface is predestined for an automated approach. For this study, we tested different DNA extraction kits using DNA-binding paramagnetic particles with regard to DNA yield and handling by a Freedom EVO(®)150 extraction robot (Tecan) equipped with a Te-MagS magnetic separator. Among others, the extraction kits tested were the ChargeSwitch(®)Forensic DNA Purification Kit (Invitrogen), the PrepFiler™Automated Forensic DNA Extraction Kit (Applied Biosystems) and NucleoMag™96 Trace (Macherey-Nagel). After an extensive test phase, we established a novel magnetic bead extraction method based upon the NucleoMag™ extraction kit (Macherey-Nagel). The new method is readily automatable and produces high yields of DNA from different sample types (blood, saliva, sperm, contact stains) on various substrates (filter paper, swabs, cigarette butts) with no evidence of a loss of magnetic beads or sample cross-contamination.
LKA35 (DNA-Analytik), Landeskriminalamt Hamburg, Bruno-Georges-Platz 1, 22297 Hamburg, Germany. lka35@polizei.hamburg.de
Sebastian
S
Witt
Jan
J
Neumann
Holger
H
Zierdt
Gabriella
G
Gébel
Christiane
C
Röscheisen
D001331:Automation; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D053803:Forensic Genetics; D006801:Humans; D008280:Magnetics; D018895:Microsatellite Repeats / Q000235:genetics
D004247:DNA