is there any benefits if I make Taq polymerase heat resistant at 100 C°? (not only...

is there any benefits if I make Taq polymerase heat resistant at 100 C°? (not only 94 C°)

Expert Solution

If Taq polymerase is made heat resistant at 100 C° rather than 940C, it gives more stability to the enzyme.

Taq DNA polymerase was isolated from a geothermal spring in Yellowstone National Park is the most well known example of biotechnological application of extremophiles and their biomolecules. Taq DNA Polymerase has a temperature optimum of 720C and survives at prolonged exposure to temperatures as high as 96oC and is active after each denaturation steps.

Polymerase Chain Reaction (PCR) consists of three defined sets of times and temperatures with three steps

1. Denaturation

2. Annealing

3. Extension

Denaturation require high temperature (96oC) to denature the DNA prior to primer annealing and during the steps of annealing and also during extension process to prevent secondary structure formation. Taq DNA polymerase are active at high temperatures, but is not quite stable at 95°C. The half life of Taq polymerase decreases with the increase of temperature above 90oC.

TEMPERATURE(IN CELSIUS)	HALF LIFE
92.5	2 Hours
95	40 minutes
97.5	9 minutes

Hence increasing the Heat resistance to 1000C will give more stability to Taq polymerase enzyme increasing its half life.

gladiator answered 2 years ago

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