After a ligation of a vector and insert, be able to draw a restriction map of th

Question

After a ligation of a vector and insert, be able to draw a restriction map of the plasmid (as shown in the diagram below). If provided with a plasmid restriction map, be able to predict the restriction pattern (number and size of gel bands) after digestion with a given restriction enzyme. Example Question: In an attempt to clone the lacZ gene, you ligate your lacZ gene insert (3 kb) into a commercially vector (4 kb long) between two EcoRI cut sites. The desired recombinant plasmid is diagramed below showing one vector (thin line) and one insert (thick line). All of the cut sites for the restriction enzymes EcoRI and PstI and the approximate distance between these cut sites are indicated. Using restriction enzymes and gel electrophoresis, what restriction pattern would you expect with an EcoRI digest? A PstI digest? An EcoRI and PstI double digest? A = 3 kb B = 1 kb C = 3 kb

Explanation / Answer

As the recombinant vector is now of 7Kb. So when cut with EcoRI the fragments will be - 3 Kb and 4Kb.

With PstI digest, we will get linearised vector of 7 Kb.

With both EcoRI and PstI, - 3Kb, 1Kb and 3Kb.

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