Applied Biosystems™ PrepFiler™ Forensic DNA Extraction Kit (Manual and Semi-automated via AutoMate Express™)

The PrepFiler™ Forensic DNA Extraction Kits allow for optimal genomic DNA isolation and purification from forensic samples through a bind-wash-elute-based technique that can be performed manually or robotically using the Applied Biosystems AutoMate Express™ Forensic DNA Extraction System. The extraction kits come in two formats: the standard kit used for common case type samples, like bodily fluid swabs or stains, and a BTA kit for more challenging evidence sample types that can be submitted for analysis, like bones, teeth, and adhesive-type samples. Both forms of extraction, manual and semi-automated, require an initial manual incubation step using a lysis buffer to release the DNA into solution. If following the semi-automated protocol, the lysate can be purified and eluted on the AutoMate Express™. After lysis, the DNA binds to magnetic beads in the presence of a chaotropic salt and is washed multiple times with an ethanol-based wash buffer to purify the sample and remove potential PCR inhibitors. After removing the wash liquid, elution buffer is added to the tube containing the DNA-bound magnetic beads and heated, which disrupts the bonding between the DNA and beads. The DNA is then concentrated in the final tube and can be moved forward through the DNA analysis workflow. This chapter describes a manual DNA isolation method and the extraction procedures following both manual and robotic methods using the PrepFiler™ chemistries in conjunction with the AutoMate Express™ Forensic DNA Extraction System.