What does hot start PCR do?|How can i speed my PCR?|

                           

                              HOT-Start PCR

Ø Hot start PCR is to use to minimize unwanted DNA product.

Ø Major disadvantages is PCR production of unwanted DNA amplification.

Ø Second step in addition to primer and tag polymerase. We add specific antibodies to back to polymerase from annealing.

Ø When temperature raises from amplification 720⁰c the specific antibodies detaches from tag polymerase and amplification being with great specificity.

Ø Modification of polymerase normally we add some antibody that will coat and being tag polymerase in normal room temperature is inactive them in room temperature antibody will properly function. So that tag polymerase can not initiate the polymerase chain reaction.

Ø As we start increase temperature at very first of the PCR reaction the temperature can 95⁰c that is call HOT-start PCR.

Ø Because the HOT-start PCR higher temperature from beginning.

Ø Then that will inactive all antibody.

Ø Antibody will be degraded so that tag polymerase. Will free so that the elongate the primer properly. 

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