Practice Problem 99 Your group is a part of a research team that just received a

Question

Practice Problem 99 Your group is a part of a research team that just received a grant from 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 p5 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 H H 0 kb 2 kb 5 kb 8 kb 10 kb 11 kb PCR product EcoRI H HindIII N Notl S The plasmid into which you want to clone the p53 gene is shown below. 10 kb B (BamHI X (Xbal) N (Not) V H (Hindlll) E (EcoRI) Submit My Answers Give Up Correct 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 Part B fter cutting the PCR product with HindlII and Notl, how many pieces of DNA will you have? Enter your answer as a whole number Submit My Answers Give Up Incorrect; Try Again; 4 attempts remaining The PCR product was only cut with Hindll and Notl. It was not cut with any of the other restriction enzymes. In how many locations do these two enzymes cut the PCR product?

Explanation / Answer

Part B- The PCR product is cut with Hind2 and Not1 which are represent by H and N. now we look on the given gene first N at 1kb cut into two fragments now the right side fragment which is 10 kb in size cut by H. H cut at the end of right side at 11 kb mark and 2nd cut at 10 kb mark so total fragments we got 3

These fragments are 0 kb to 1kb by N , 1kb to 10kb by H and 10kb to 11 kb by H.

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