füüsikalises distantsis fluorokroomi ja vaigistaja (quencher) vahel pärast proovide hübridisatsiooni. FRET (fluoresence resonance energy transfere) tehnika -Iga SNP vajab spetsiaalpraimereid -Kõige efektiivsemad väheste SNP-de ja suure hulga proovide puhul Introduction to TaqMan® probes TaqMan® probes are dual labeled hydrolysis probes and are a registered trademark of the Roche Molecular Systems, Inc. TaqMan® probes utilize the 5' exonuclease activity of the enzyme Taq Polymerase for measuring the amount of target sequences in the samples. TaqMan® probes consist of a 18- 22 bp oligonucleotide probe which is labeled with a reporter fluorophore at the 5' end and a quencher fluorophore at the 3' end. TaqMan® probe functioning While carrying out a TaqMan® experiment, a fluorogenic probe, complementary to the target sequence is added to the PCR reaction mixture
The (Förster 1948; Clegg 1992), and a change in initial DNA concentration of the sample the distance between them is used to produce can then be determined by interpolation the RTi-PCR signals. The hydrolysis probes of the resulting CT value in a linear standard such as TaqMan® probes and TaqMan® MGB curve of serially diluted, known-amount probes are cleaved by the 5′-3′ exonuclease standards. activity of several DNA polymerases during the primers’ elongation phase (Holland et al. Other DNA Techniques 1991), yielding a real-time, measurable fluo- rescence emission directly proportional to the Other modifications and techniques based on concentration of the target sequence. In con- DNA detection are continuously appearing in
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