the National Institutes of Health to study cancer As a first

the National Institutes of Health to study cancer. As a first step, you decide to clone p53, a human gene that is mutant in many types of cancer cells, Below is a diagram of the DNA product of a PCR amplification of the region of the human genome containing p53. The white region represents the coding region of the gene. The letters above the DNA represent the locations of specific restriction enzyme cut sites. The numbers below each restriction site represent the number of kilobases (kb) from the first restriction site (at 0 kb). Each of the restriction enzymes creates unique sticky ends. The plasmid into which you want to clone the p53 gene is shown below. If the PCR product is cut with N and H, unique sticky ends are generated on both sides of the complete p53 gene. N and H sites are both present in the multiple cloning site of the plasmid: therefore, the p53 gene could be ligated into this region of the plasmid without interfering with any of the essential features of the plasmid. After cutting the PCR product with HindIII and Non, how many pieces of DNA will you have? Enter your answer as a whole number. When the linear PCR product is cut with the two enzymes, N and H, three pieces of DNA result. The middle piece of DNA-the one between the N site and the first H site-contains the entire p53 gene. After cutting the plasmid with HindIII and Non, how many pieces of DNA will you have? Enter your answer as a whole number. This question will be shown after you complete previous question(s).

Solution

Answers: (Part B has already been answered correctly)

Part C: 2 fragments

After cutting the plasmid with NotI, we shall get a linearized DNA of 14kb. Again after digestion with HindIII, we shall get 2 fragments. The region between NotI and HindIII sites is 20 bases. So the size of the fragments would be 20 bases and 13.980 kb.