Comparative study on the effects of reduced PCR reaction volumes and increased cycle number, on the sensitivity and the stochastic threshold of the AmpFlSTR Identifiler® Plus kit

Abstract

The purpose of this study is to compare the performance of reduced volume PCR reactions with the Identifiler Plus kit and to investigate the influence on the sensitivity and stochastic occurrences under standard and increased thermal cycling conditions.Among the three volumes, sensitivity and stochastic effects appeared to be unchanged within each cycling condition. Under the increased cycle condition, the sensitivity was higher than under standard cycling conditions but there was also a consequent increase in background noise and stochastic threshold. The limitation of reducing volumes appears to be mainly due to the challenge of small volume manipulation. A 10μL final volume proved to be both efficient and economical while simultaneously significantly decreasing the amount of evidence consumed.