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The main problem with enzymes is that they are very sensitive to their surroundings. For enzymes to function properly the temperature must be kept down (usually below 45oC). The pH also needs to be kept within carefully monitored limits which suit the enzyme. It costs money to control these conditions.ØEnzymes are very sensitive to their surroundings. For enzymes to function properly the temperature must be kept down (usually below 45oC). The pH also needs to be kept within carefully monitored limits which suit the enzyme. It costs money to control these conditions.
ØSmall changes in these conditions will make them denatured
ØEnzymes require a lot of energy
ØPeople can develop Allergies to enzymes such as biological washing powder
ØThey are expensive to produce
ØContamination of the enzyme with other substances can affect the rate of reaction.
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How are large DNA molecules cut up?
by using restriction enzymes
What are the advantages of using two restriction enzymes in clonning over one?
· For the DNA to be inserted into the vector the two ends must be different. Using two restriction enzymes ensures that there are two different sticky ends to the DNA and thus insuring that the DNA is inserted in the correct orientation. · Another reason for this is that using two enzymes you can tell which direction the enzyme is working. · One major advantage of using two restriction enzymes for cloning is that you can control the orientation that your DNA fragment is inserted into the vector (especially useful if you're making a construct to express protein).
