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Which of the following is not a step involved in discontinuous assay?

(a) Taking sample from the reaction mixture at known time

(b) Stopping the reaction by inactivating the enzyme

(c) Measure the convenient change in absorbance

(d) Measuring the amount of product formed

The question was posed to me during an interview.

Question is taken from Continuous, Discontinuous and Coupled Assay topic in section Enzyme Assay & Enzyme Catalysis of Enzyme Technology

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Best answer
The correct option is (c) Measure the convenient change in absorbance

The best explanation: “Measure the convenient change in absorbance.” This step is involved in continuous assay, and not discontinuous, which has “stop and sample procedure”. The steps involved in discontinuous assay are as follows:

• Taking sample from the reaction mixture at known time after addition of enzyme.

• Stopping the reaction by addition of acid to inactivate the enzyme.

• Measuring the amount of product formed.

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