Imagine that you collected your data above, threw away the gel, and re-ran PCR again on the same samples, but using a different set of primers. How will the 2nd gel differ from the first since you're using a different set of primers?
You set up 5 samples that contained a range of sucrase concentrations but the same amount of
substrate (sucrose).
- The pH was kept constant in all tubes by using the same amount of buffer.
- The temperature was kept constant in all tubes (room temp)
- A negative control was used (without the substrate sucrose) to prove that this reaction is due to enzyme activity. It is important to establish this, because sucrose can break down on its own to yield glucose and fructose but this reaction would take much longer without enzyme.
- There was no quantitative data, but a color key was used to distinguish between maximum, medium and low enzyme activity.
- By graphing the qualitative data, you observed that the reaction increases as the enzyme concentration is increased. This means, the more enzyme, the more sucrose can be broken down, the more products can be formed.
- However this effect can be observed up to a limit.
As stated, the results as evidenced in the experiment can only be observed up to a limit. Why?