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Primer extension may be hindered due to the presence of template structures at extension temperature. To avoid all such regions, where the template may fold upon itself, the program utilizes a proprietary algorithm to check for possible secondary structures within the template at a folding temperature you specify. The regions involved in the formation of a secondary structure are underlined in the sequence view and are avoided while designing primers.
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"The primers that your program designed pretty much all worked to produce working taxa-specific primers with great efficiencies and R2 values on a standard dilution series to be used for qPCR. I really did enjoy the trial of AlleleID, what a great program!" -Mr. Ben Sutherland, University of Victoria, Canada.
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Addition of MagPlex TAGs
The program now also supports a database of MagPlex TAGs along with the MicroPlex xTAGs (formerly known as FlexMAP TAGs) for automatic addition of appropriate tags to each ASPE primer. The tags are so chosen that they minimize dimerization and do not fold back on the oligos they are attached. In addition, a user can override the program's recommendation and select a tag on their own. PrimerPlex alerts the user if the tags are not unique or if the tag chosen is incompatible with the oligo.
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Exploring target-specific primer extension in combination with a bead-based suspension array for multiplexed detection and typing using Streptococcus suis as a model pathogen
Fimme J. der Wal, René P. Achterberg, Conny Solt-Smits, Jan H. W. Bergervoet, Marjanne de Weerdt, Henk J. Wisselink.
Journal of Veterinary Diagnostic Investigation. 2018; Jan; 30(1): 71–.
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