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Real Time PCR procedure
In a real time PCR protocol, a fluorescent reporter molecule is used to monitor the PCR as it progresses. The fluorescence emitted by the reporter molecule manifolds as the PCR product accumulates with each cycle of amplification. Based on the molecule used for the detection, the real time PCR techniques can be categorically placed under two heads:
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.
Cross Species Detection
Changes in the DNA makeup of the organisms such as single nucleotide polymorphisms, during the course of evolution, have led to the diversity in phenotypic traits. Generations after generations, inherit these changes and contribute towards the biodiversity of species. Despite all these changes, there are still stretches of DNA which remain conserved during evolution. Such conserved regions or sequences are useful in suggesting phylogenetically similar groups, identification of an organism within a group or understanding the extent of its biodiversity amongst the flora of a particular area.
Species Identification: Minimal probes with Optimal primers
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