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Taxa Specific/Cross Species Identification: Conserved Probe with Optimal Primers
Conserved Probe with Optimal Primers: AlleleID® designs a primer pair that would amplify the maximum number of sequences possible in an alignment. There are two choices available for designing primers, Unique and Optimal. For "Unique", the program designs a primer pair to amplify each sequence. Each primer is completely homologous to its template. For "Optimal', the program minimizes the number of primers required to amplify all sequences; taking advantage of the fact that primers can bind to a sequence and amplify it even if they contain a limited number of mismatched bases especially towards the 5' end of the primer.
Choices for Primer Design: There are two choices available for primer design, Unique and Optimal. When you choose Unique, the program designs a primer pair to amplify each sequence. For Optimal, the program minimizes the number of primers required to amplify all sequences. The goal is to minimize the synthesis and overall assay costs.
Real Time PCR & Microarray Primer/Probe Design for Cross Species and Taxa Specific Applications
For cross species assays, AlleleID® identifies the conserved regions to design a universal probe. This functionality is especially useful to detect a species or a taxa when designing a diagnostic assay. This functionality is also useful to help study gene expression when genome draft of the organism under study is not available.
When it is not possible to identify a significant conserved region for a set of sequences, AlleleID® attempts to design a "Mismatch Tolerant" probe. The probe may include up to the specified number of mismatched bases which are located as close as possible to the 5' end.
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.
Taxa Specific/Cross Species identification: Probes with Mismatched Bases and Optimal Primers
Minimal Probes with Optimal Primers: When the homology among the aligned sequences is very low and both the designs mentioned above (a single taxa specific probe set or probe set with mismatch tolerance) are not feasible, AlleleID® designs the minimum number of probes possible to detect all the sequences in an alignment. The program designs probes for the majority consensus first and then for the minority consensus and remaining individual sequences. This design is very useful, especially for studying phylogenetic relationships among sequences obtained from different geographical locations.
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