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To explain: The property of DNA that is influenced by the temperature in PCR.
Introduction: PCR stands for polymerase chain reaction, which is an invitro laboratory technique used for the synthesis of multiple copies of specific DNA segments of interest. This technique is carried out in a thermocycler to generate more copies of DNA.
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Explanation of Solution
The PCR technique is performed under high temperatures to amplify the DNA fragments. The intertwined nature of DNA is influenced by the temperature in PCR. The double strands of the DNA are tightly bonded together by the hydrogen bonds between the bases of each strand. These strands provide stabilization to the DNA molecule. A higher temperature is required to break the bonding and to unwind the two helices of the DNA separately.
To explain: The reason for the denaturation and annealing steps to proceed at different temperatures.
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Explanation of Solution
Denaturation is the initial step of PCR in which the double-stranded DNA template is separated into two single strands by the application of extreme temperature. It occurs at a temperature of 95°C.
Annealing is the second step in which the primers are attached or joined or annealed to the separated DNA template. This step is carried out at the temperature between 48–72°C.
The denaturation and annealing steps are carried out under different temperatures. This is because the higher temperature in the first step facilitates the denaturation of DNA strands. Such a high temperature would not facilitate the annealing of primers to the DNA strands in the second step.
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LooseLeaf Benson's Microbiological Applications Laboratory Manual-Concise Version
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