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Species Identification Assays/Cross Species Assays/Allele Identification Assays
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.
Modern Techniques for Cross Species Detection
Real time PCR and microarrays offer a rapid, sensitive, specific and quantitative tool for the diagnosis of pathogens. Diseases are not readily distinguished based on symptoms when there is more than one pathogen in the sample. Microscopic identification requires culturing on specialized media for days to weeks. Recently, real time PCR assays were successfully used in the detection of fungal pathogens such as Rhizoctonia, Pythium, and Fusarium.
Designing a cross species assay involves the use of a multiple sequence alignment tool to identify conserved stretches of DNA and the design of a hybridization probe for detecting it. Recently there have been advances in micro-organism identification at the taxonomical level using molecular level identification techniques such as real time PCR and microarrays.
AlleleID® Provides Several Design Options for Designing Taxa Specific/Cross Species Assay
Conserved Probes with Conserved Primers: AlleleID® identifies conserved regions by aligning sequences and then designs a probe within them. This enables identification of any sequence in the set with a single probe. It also designs primer pairs in the conserved regions flanking the probes. As a result, a single primer pair can amplify all the sequences of a group or a taxa.
Also included is the "Minimal Set" option that helps you design minimum number of probe sets that uniquely identify a sequence reducing the overall assay cost.
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