Rapid direct PCR for forensic genotyping in under 25 min

Abstract

In this paper, a rapid thermal cycling procedure is combined with a direct amplification from a paper punch, permitting a high-speed amplification of a 7-locus multiplex that requires no extraction step. When coupled with a short 1.8 cm microfluidic electrophoresis system, the entire procedure from paper punch to genotype can be completed in under 25 min. The paper describes selection and optimization of enzyme, direct amplification conditions, the reproducibility of the procedure, and concordance with standard forensic genotyping methods. The procedure utilizes a small high-speed thermal cycler and microfluidic device along with a small laptop and is highly portable. Overall, this technique should provide a useful and reliable procedure for rapid determination of identity of individuals retained at checkpoints as well as a quick method for preliminary identification of individuals at remote locations following mass disasters.