Ploxpflpneo plasmid schematic questions 1 what are

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ploxPFlpneo plasmid schematic
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Questions: 1. What are some potential reasons as to why the 5’ arm was so difficult to clone into the SwaI restriction site (before the SwaI polylinker was introduced)? 2. What is the most likely reason why the 5’ arm must be cloned into this plasmid before the 3’ arm? 3. Which pair(s) of restriction sites (if any) can be ligated to each other? Only consider the restriction sites provided above. 4. Why was ApaI included into the SwaI polylinker? Hint: what will you get if you digest this plasmid with ApaI and ran the result on an agarose gel as opposed to any other restriction site? 5. What would you do to test whether the SwaI polylinker had been successfully inserted into the ploxPFlpneo plasmid? 6. Diagram what you would see on an agarose gel if you digested this plasmid with ApaI. Now diagram what you would see if you digested this plasmid at any other restriction site. Assume that all fragments (Exon 4, 5’ arm, and 3’ arm) have all been inserted and round to the nearest tenth of a Kb. 7. Locate and diagram all the restriction sites in the SwaI polylinker. What do you notice?
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