Describe the steps of PCR in terms of the specific temperatures and what happens

Question

Describe the steps of PCR in terms of the specific temperatures and what happens at each stage of each cycle. Be able to describe what would happen (in terms of enzymatic activity and amount of PCR product produced) if you decreased OR increased any of the conditions in any of the steps. Describe he function of each regent and what role it plays in making a PCR proceed correctly. Describe the steps of PCR in terms of the specific temperatures and what happens at each stage of each cycle. Be able to describe what would happen (in terms of enzymatic activity and amount of PCR product produced) if you decreased OR increased any of the conditions in any of the steps. Describe he function of each regent and what role it plays in making a PCR proceed correctly.

Explanation / Answer

PCR is a technique used to amplify DNA content. the steps in PCR are-

1. DENATURATION- this is done on 90-94 degree C where the DNA denature and double helix gets separated. it is a specific temparature as DNA melting point.

2. ANNEALING- the temparature is lowered to 6 0-64 degree C where DNA polymerase enzyme of Thermus aquaticus is added along with oligonucleotide primers.

3. Then the temparature lowered to 40-45 degree where synthesis or copy of the DNA occur which amplify the DNA content.

the all step carried out in thermal containers at specific temparature as each step is highly specific. so if there is any increase or decrease in temparature the above any of the step will not occur properly as PCR is a 3 step chain reaction we may not get the amplified PCR products upon changing the temparature at any step.

Taq polymerase is used as it is stable in high temparature.

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