HTself is a web-based bioinformatics tool designed to deal with the classification of differential gene expression in low replication microarray studies. It is based on a statistical test that uses self-self experiments to derive intensity-dependent cutoffs. We developed an extension of HTself, originally released in 2005, by calculating P values instead of using a fixed acceptance level α. As before, the statistic used to compute single-spot P values is obtained from the Gaussian kernel density estimator method applied to self-self data. Different spots corresponding to the same biological gene (replicas) give rise to a set of independent P values that can be combined by well-known statistical methods. The combined P value can be used to decide whether a gene can be considered differentially expressed or not. HTself2 is a new version of HTself that uses P values combination. It is implemented as a user-friendly desktop application to help laboratories without a bioinformatics infrastructure.