This article reports on a rapid analysis procedure for use with a small set of rapidly mutating Y chromosomal short tandem repeat (Y-STR) loci that combines both rapid polymerase chain reaction (PCR) and microfluidic separation elements.
The procedure involves a high-speed polymerase and a rapid-cycling protocol to permit PCR amplification in 16 minutes. The resultant amplified sample is next analyzed using a short 1.8-cm microfluidic electrophoresis system that permits a four-locus Y-STR genotype to be produced in 80 seconds. The entire procedure takes less than 35 minutes from sample collection to result. This paper describes the rapid amplification protocol as well as studies of the reproducibility and sensitivity of the procedure and its optimization. The amplification process uses a small high-speed thermocycler, micro fluidic device and compact laptop, making it portable and potentially useful for rapid, inexpensive on-site genotyping. The four loci used for the multiplex were useful for rapid, inexpensive on-site genotyping. The four loci used for the multiplex were selected due to their rapid mutation rates and should prove useful in preliminary screening of samples and suspects. Overall, this technique provides a method for rapid sample screening of suspects and crime-scene samples in forensic casework. 1 figure (Publisher abstract modified)