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You Are Attempting to Generate a Physical Map of the Bacteriophage

Question 19

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You are attempting to generate a physical map of the bacteriophage λ\lambda genome using restriction enzyme digestions. You digest the phage DNA with EcoRI or NcoI or both, and observe the fragment sizes listed below. You know that bacteriophage λ\lambda has a linear, not circular, genome.  EcoRI  NcoI  EcoRI/NcoI 21.2 kb19.3 kb19.3 kb7.4 kb16.4 kb7.4 kb5.8 kb4.6 kb5.1 kb5.7 kb4.2 kb3.9 kb4.9 kb4.0 kb3.5 kb3.5 kb2.7 kb2.2 kb1.9 kb1.8 kb0.7 kb\begin{array} { l l l } { \text { EcoRI } } & { \text { NcoI } } & { \text { EcoRI/NcoI } } \\\hline { 21.2 \mathrm {~kb} }&{ 19.3 \mathrm {~kb} } &{ 19.3 \mathrm {~kb} } \\7.4 \mathrm {~kb} & 16.4 \mathrm {~kb} & 7.4 \mathrm {~kb} \\5.8 \mathrm {~kb} & 4.6 \mathrm {~kb} & 5.1 \mathrm {~kb} \\5.7 \mathrm {~kb} & 4.2 \mathrm {~kb} & 3.9 \mathrm {~kb} \\4.9 \mathrm {~kb} & 4.0 \mathrm {~kb} & 3.5 \mathrm {~kb} \\3.5 \mathrm {~kb} & & 2.7 \mathrm {~kb} \\& & 2.2 \mathrm {~kb} \\& & 1.9 \mathrm {~kb} \\& & 1.8 \mathrm {~kb} \\& & 0.7 \mathrm {~kb}\end{array} a. How many times does the EcoRI enzyme cut the λ\lambda DNA? How many times does the NcoI enzyme cut the λ\lambda DNA?
b. Use the fragment size data to generate a physical map of the bacteriophage λ\lambda genome, indicating the order and relative positions of the restriction enzyme recognition sites.

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a. To determine how many times the EcoRI...

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