Why are protease inhibitors used during homogenization? To protect protein samples from being degraded by proteases To protect DNA from being degraded by proteases To protect RNA from being degraded by proteases
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Which of the following techniques are useful to prevent RNA degradation during an experiment? A) Use a buffer to maintain alkaline conditions (pH = 13) to stop any cleaving enzymes from working. B) Use a buffer containing cleaving agent(s) to stop any cleaving enzymes from working. C) Keep samples on ice. Use a buffer to maintain extremely acidic conditions (pH = 0.5) to stop any cleaving enzymes from working. D) Incubate samples at high temperatures to stop any cleaving enzymes from working.
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Step 3: Using protease to break down cellular proteins The most prevalent class of molecules that would interfere with the precipitation of pure DNA is protein. We can easily get rid of protein without damaging the DNA by using a specific enzyme that digests proteins, called a protease. Protease breaks the peptide bonds between the amino acids of proteins. By destroying all the proteins you will also eliminate DNases, enzymes that digest DNA (because enzymes are proteins). FOCUS QUESTIONS 8. What proteins might be associated with DNA in the cell? 9. The protease used in this procedure functions best at 50°C. Would you expect this enzyme to be isolated from E. coli bacteria? Explain your answer. Hint: Where does E. coli live? 10. Meat tenderizer is often used to tenderize tough pieces of meat, like steak. Knowing that steak is made of protein-rich muscle tissue from cows, can you think of an explanation for how meat tenderizer works?
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The main purpose of adding EDTA to cell lysate is to? Select one: a. Protect genomic material from degradation b. Enhance the precipitation of the genomic material c. Reduce the amount of impurities co-precipitated with RNA d. To increase the quantity of the extracted material e. To enhance the binding of DNA molecules to silica membrane
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