Dialyzed saliva (from a source entirely different than the one used in your expe

Question

Dialyzed saliva (from a source entirely different than the one used in your experiment) led to an achromic point of 5 minutes while undialysed saliva led to one of 8 minutes? Furthermore, addition of either NaCI or KCI to the dialyzed saliva led to an achromic point of 10 minutes. Dilution and assay conditions were otherwise the same as what you did in the lab. When setting up an enzyme reaction, it is good practice to prepare a non-enzymatic control (to check on activity) and pay particular attention to concentrations and amounts of components. Use the following information to complete the table. 10 X React3 buffer DNA (1.0 mg mL^-1) EcoRI (2.0 U mu L^-1) Distilled Water EcoRI works best at 1X use 1.0 mu g use 2.0 U enzyme per mu g DNA adjusted to control total volume How would your results from #3 be affected if you failed to mix the reaction mixtures to homogeneity prior to incubation at 37 degree C for 30 minutes?

Explanation / Answer

3.

4. when we mix the reaction mixture to homogeneity all the component of the reaction are mixed properly and we get an even mixture of the reaction mix. if we failed to mix the reaction mixture to homogeneity before incubating then we might not get the properly digested DNA or DNA could not digested at all.

- enzyme +enzyme water 24 25 10X buffer 3 3 DNA 1 1 EcoR1 none 1 Total 30 30

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