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Annealing Time and Temperature

Molecular Diagnostics
Edited by: Jim Huggett and Justin O'Grady
The application of molecular technology in clinical diagnosis in two key diagnostic areas: cancer and infectious diseases.
I would highly recommend this book (Doodys) read more ...
Real-Time PCR
Edited by: Nick A. Saunders and Martin A. Lee
Provides both the novice and experienced user with an invaluable reference to a wide-range of real-time PCR technologies and applications and supplies detailed technical insights into the underlying principles, methods and practice of real-time PCR.
"an invaluable reference" (Doodys); "wide range of real time PCR technologies" (Food Sci Technol Abs) read more ...

Annealing Time

An annealing time of 30-45 seconds is commonly used in PCR reactions. Increase in annealing time up o 2-3 minutes did not appreciably influence the outcome of the PCR reactions. However, as the polymerase has some reduced activity between 45 and 65o C (interval in which most annealing temperature are chosen), longer annealing times may increase the likelihood of unspecific amplification products.

Annealing Temperature

Annealing temperature is one of the most important parameters that need adjustment in the PCR reaction. Moreover, the flexibility of this parameter allows optimization of the reaction in the presence of variable amounts of other ingredients (especially template DNA).