AlleleID® designs two types of microarray probes that we call junction probes and exon probes, giving the user complete control over the design parameters.
AlleleID® detects the alternative splicing events by designing probes for each junction and/or for each splice variant created.
AlleleID® designs exon probes. Such probes are important to confirm the presence of an exon in the variant.
Technical resource on Gene Splicing.
The coding and non-coding fragments of the gene can be arranged in different ways. When this produces different m-RNA sequences from the same parent gene, the phenomenon is known as Alternative Splicing. The biological consequences of this process can be severe as the same gene leads to formation of different proteins which may be functional, non-functional or malfunctioning. The varied forms of the splicing events are known as splice variants. And as outlines above, AlleleID® could be used for splice variant detection using microarray experiments.
AlleleID® aligns sequences using ClustalW and analyzes conserved and species specific regions. You can then use the program for real time PCR primer design (SYBR® Green primer design included) and dual labeled probe design (TaqMan® probes, TaqMan® MGB probes and molecular beacons). These assays are designed to detect only the strain (strain detection) or species of interest from the mix.
Highly specific oligos are designed by avoiding regions of significant homologies found by automatically interpreting BLAST search results. Real time PCR primer & probe efficiency is enhanced by avoiding template secondary structures. "Minimal Set", one of the most innovative features in the program, helps design the fewest number of allele specific oligonucleotide primers and dual labeled probes that uniquely identify each of the desired species/strain/taxa from the mix, lowering assay costs. For taxa or cross species assays, this feature is especially useful when the group or taxa is highly dissimilar. For a partial set of pre-designed, proven set of primers, AlleleID® can design compatible primers and probes for the rest of sequences for species identification or taxa specific assays.