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Although these double-stranded DNA-binding dyes provide the simplest and cheapest option for real time PCR, the principal drawback to intercalation based detection of PCR product accumulation is that both specific and nonspecific products generate **.
Molecular based techniques use species-specific probes, designed for specific regions of the target gene, for the detection and identification of bacteria. Microarrays can rapidly detect and identify multiple pathogens simultaneously in complex backgrounds.
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.
What is Alternative 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.
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