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Question: 8. Using the maps provided to you, develop a cloning strategy to insert the Cycle 3 GFP gene, now…

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Show transcribed image text 8. Using the maps provided to you, develop a cloning strategy to insert the Cycle 3 GFP gene, now in the pBlueScript vector (seen in the recombined plasmid in Figure 1 on the next page), into the expression vector pCR-XL-TOPO (Figure 2 on the next page). Assume that your insert is in the correct orientation in the pBlueScript vector. Then answer the questions and do the problems below. a) What restriction enzyme(s) will you use to cut and paste your gene of interest (Cycle3 GFP) into the pCR-XL-TOPO plasmid? b) Will you be doing a double or single digest and why? Will you need to use CIP in your digest? If so, why? If not, why not? c) Draw a map of your new recombinant pCR-XL-TOPO/GFP mammalian expression vector

8. Using the maps provided to you, develop a cloning strategy to insert the Cycle 3 GFP gene, now in the pBlueScript vector (seen in the recombined plasmid in Figure 1 on the next page), into the expression vector pCR-XL-TOPO (Figure 2 on the next page). Assume that your insert is in the correct orientation in the pBlueScript vector. Then answer the questions and do the problems below. a) What restriction enzyme(s) will you use to cut and paste your gene of interest (Cycle3 GFP) into the pCR-XL-TOPO plasmid? b) Will you be doing a double or single digest and why? Will you need to use CIP in your digest? If so, why? If not, why not? c) Draw a map of your new recombinant pCR-XL-TOPO/GFP mammalian expression vector