ProbeMaker: an extensible framework for design of sets of oligonucleotide probes

ProbeMaker is a software framework for computer-assisted design and analysis of sets of oligonucleotide probe sequences. The tool assists in the design of probes for sets of target sequences, incorporating sequence motifs for purposes such as amplification, visualization, or identification. An extension system allows the framework to be equipped with application-specific components for evaluation of probe sequences, and provides the possibility to include support for importing sequence data from a variety of file formats.ProbeMaker is a suitable tool for many different oligonucleotide design and analysis tasks, including the design of probe sets for various types of parallel genetic analyses, experimental validation of design parameters, and in silico testing of probe sequence evaluation algorithms.Increasing numbers of methods are being developed for parallel nucleic acid analyses for different purposes. Many of these methods employ sets of oligonucleotide probes or probe pairs that hybridize to the sequences targeted for analysis, allowing the probe sequences to be acted upon by one or more enzymes, creating new molecular species that reflect the presence or nature of the different target sequences. The reaction products generally contain identifying sequences or other features that allow the separation of signals originating from different targets. This is the case in methods such as the multiplex oligonucleotide ligation assay (OLA) [1], the multiplex ligation-dependent probe amplification assay (MLPA) [2], the RNA- and cDNA-mediated annealing, selection, extension and ligation assays (RASL, DASL) [3,4], the GoldenGate genotyping assay [5], multiplex minisequencing [6], and the padlock or molecular inversion probe assay [7,8]. The latter method has been used to genotype more than 10,000 single nucleotide polymorphisms (SNPs) in multiplex. Another method that utilizes sets of oligonucleotide probes for multiplex processing of nucleic acid molecules is the selecto