Enhancement of DNA recovery in rapid DNA technology: A novel pre-extraction protocol

Forensic laboratories aim to shorten the time needed to produce a DNA profile without compromising its quality. During the past decade, Rapid DNA technologies emerged, introducing fast generation of DNA profiles. Despite remarkable progress in rapidly processing high-DNA-content specimens within 90 min, current methods still face challenges. These limitations affect the efficiency of DNA extraction and the quality of the resulting profiles, depending on the item types processed. We present an optimized pre-extraction protocol that introduces a 12-minute preparatory phase of thermal agitation to enhance low-template DNA recovery, using the latest RapidHit™ cartridges IntelPlus (Thermo Fisher Scientific). Incorporating thermal agitation significantly improved profile quality in diverse forensic items. Pre-extracted profiles had more alleles called, higher allele peak heights and a finer balance of peak height ratios (PHR). Additionally, the pre-extracted samples were less degraded, and DNA yield was higher according to internal quantity control markers.