Following information is from the instruction of Q5 DNA polymerase: One unit of enzyme will incorporate...

Following information is from the instruction of Q5 DNA polymerase: One unit of enzyme will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 74°C. The temperature of denaturing should be ( ) and the temperature of extension should be ( )

		94℃ to 98℃ for both.
		94℃ to 98℃ for denaturing, 74℃ for extension.
		72℃ for denaturing, 50℃ to 65℃ for extension.
		94℃ to 98℃ for denaturing, 50℃ to 65℃ for extension.

Expert Solution

The correct answer is 2nd option, i.e. 94℃ to 98℃ for denaturing, 74℃ for extension.

Reason : This initial denaturing temperature is preferably set between 94-98° C for most samples. The reason behind this is the fact that this temperature range is found to be optimal for denaturing the DNA sample. Temperature higher than this range will surely damage the DNA bases, and lower temperatures will prevent proper denaturing as the two separated strands will bind together again. They won't properly separate at lower temperatures. So experimentally 94-98°C temperature range was found optimal. That's why other options are incorrect.

Similarly, for extension step 74°C was found to be optimal. This step is performed by DNA polymerase enzyme. Every enzyme has its own optimal temperature. Below that temperature, Enzyme activity is less. On the other hand, if the temperature is higher then the Enzyme will get denatured itself and become deactivated. So experimentally 74°C was found optimal for extension step in most DNA samples. That's why other options are wrong.

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gladiator answered 1 year ago

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