Standard PCR – Product Selection Guide - Jena Bioscience
Standard PCR – Product Selection Guide - Jena Bioscience
Standard PCR – Product Selection Guide - Jena Bioscience
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10<br />
Real-Time <strong>PCR</strong><br />
<strong>Jena</strong> <strong>Bioscience</strong> q<strong>PCR</strong> reagents provide exceptional sensitivity and accuracy<br />
Principle of real-time DNA quantifi cation using dual labeled fl uorescent probes<br />
1<br />
A quantitative real-time <strong>PCR</strong> assay requires qPol, the dual<br />
labeled fl uorescent probe, dNTPs, primers and template<br />
DNA. The proximity of fl uorophore and quencher prevents<br />
the reporter dye on the probe from fl uorescing.<br />
2<br />
The dual labeled fl uorescent probe and the <strong>PCR</strong> primers<br />
bind to their target sequences during the annealing<br />
step.<br />
3<br />
During the <strong>PCR</strong> extension step, the qPol enzyme extends<br />
the primer.<br />
4<br />
When the enzyme reaches the probe, its 5’➞ 3’ exonuclease<br />
activity cleaves the fl uorophore from the probe.<br />
The fl uorophore is released and its fl uorescence signal<br />
is detected.<br />
5<br />
After complete extension of the amplicon the next <strong>PCR</strong><br />
amplifi cation cycle is run. The measured fl uorescence<br />
increase is proportional to the amount of accumulated<br />
<strong>PCR</strong> product.
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