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Real Time PCR - Some Basic Principles

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Temperature Cycles in Real Time PCR

Figure 6. A light cycler PCR machine used in a laboratory.

Now that we have a working definition of real time PCR, we will learn how this technique actually works. The procedure of real time PCR is very similar to conventional PCR (see Polymerase Chain Reaction ). Both real time and conventional PCR require heating and cooling cycles throughout the reaction. The heating cycles denature the DNA templates and the cooling cycles allow for primers to anneal to their target. In some cases of real time PCR, a specially labeled probe also binds during the cooling cycle. The key difference in the two types of PCR systems lies in how the amplified DNA sequences are detected. Figure six shows an example of equipment used in real time PCR experiments.


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