1. What is the importance of sticky note?

- Sticky note gives specificity when rejoining different pieces of DNA.

2. The specific digestion reaction temperature and minute on the incubation phase?
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3. What is the component of the enzyme that allows it sink at bottom of the tube?
- The component that allows the enzyme to sink at the bottom of the tube is the glycerol.
Because the enzyme is stored on it.
4. What is the type of nucleases as we have previously discussed during lecture?
- The type of nuclease that we previously discuss are the Endonuclease and Exonuclease.
5. What is the difference of these two nucleases?
- The nucleotide is removed one at a time by Exonuclease at the end of the DNA molecule,
Endonuclease on the other hand hydrolyzes the internal bonds within a polynucleotide
chain.
6. How will you assure that the digest has been successful?
- We can assure that the digest is successful if there are only two bands seen in the plasmid
sample. The extracted plasmid backbone band should be higher and the PCR product
band after digest, then we can now mix these together for ligation.
7. At what volts are sample run during electrophoresis?
- The sample run at 110 volts during the electrophoresis for about 20 to 30 minutes.
8. What are the two-ideal band of digest completion?
- The backbone and the top band old gene at the bottom are the ideal band.
9. If the plasmid only cuts at one site, what band would be visible in the gel?
- The Linear piece of DNA which has the same length as the circular parental plasmid would
still remain visible. This happen because the linear DNA migrates through the gel slower
than circular DNA and appears at the higher position than the parental plasmid.
10. What is the trouble shooting that we may do if we experience a problem during the
process?
- If we experience any problem during the process it may be because the enzyme is in a bad
condition, we can order a new one or we increase the incubation time for the digest but not
longer than 4 hours.
11. Explain what is star activity.
- Star activity is the result of the incubation time that exceeds beyond 4 hours since
endonuclease of the enzyme will cause them to cut its position rather than on specific site
which can lead to a different band pattern.