6. Suppose you had a brain cramp and oriented the agarose gel in the reverse dir
ID: 141971 • Letter: 6
Question
6. Suppose you had a brain cramp and oriented the agarose gel in the reverse direction in your gel chamber. What would happen to the DNA you loaded into the well if you applied a current to the gel system with a backwards gel? 12] (think about the charge of DNA and how electrophoresis works) 7. We used 37 C as the incubation temperature for our restriction enzymes. If we had used 45°C, we would have encouraged so-called 'star activity. What does star activity mean in the context of restriction enzymes? [2]Explanation / Answer
1) DNA has a net negative charge mainly because of its phosphate backbone. It moves to the positively charged anode during electrophoresis. But when the gel is placed in the wrong direction, the negatively charged DNA will not move, and the separation will not occur.
2) A restriction enzyme is an enzyme that cleaves DNA at specific regions known as the restriction sites. This gives rise to fragments of DNA. These restriction enzymes are specific to their restriction sites only when they work in the optimum/standard conditions. When there is a major change in the optimum/standard conditions provided to the restriction enzymes, they show a change in their specificity to their restriction sites. This alteration in the activity of the restriction enzyme is known as star activity.
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