Restriction mapping of a linear piece of DNA reveals the following EcoRl restric

Question

Restriction mapping of a linear piece of DNA reveals the following EcoRl restriction sites Restriction mapping of a linear piece of DNA reveals the following EcoRI restriction sites EcoRI site1 EcoRI site 2 5 kb In each of the scenarios described below, DNA is cut by EcoRI and the resulting fragments are separated by gel electrophoresis. The gel image is pictured on the right A B C D E kb Match each of the scenarios to the lane on the gel that would result after digestion and gel electrophoresis 4 kb 3.5 kb 2 kb Continued below... Digestion is performed on the linear DNA as shown above A mutation that alters EcoRI site 1 occurs in this piece of DNA.

Explanation / Answer

Digestion performed on the linear DNA would result in the generation of 2 kb, 4 kb, and 5 kb DNA fragments. Digestion of linear DNA by EcoRI at site 1 will generate a 2 kb DNA fragment and a 9 kb DNA fragment. Further digestion of DNA with EcoRI at site 2 will fragment the 9 kb DNA into 4 kb and 5 kb DNA fragments. So, lane C represents the scenario that would result after digestion is performed on linear DNA. If there is mutation in DNA that alters the EcoRI site 1, then EcoRI cannot fragment the DNA at site 1. However EcoRI fragments the DNA at site 2 generating 6 kb and 5 kb DNA fragments. So, lane A represents the scenario that would result after digestion is performed on DNA in which a mutation has altered the EcoRI site 1. If there are mutations that alter both EcoRI site 1 and EcoRI site 2, then the enzyme cannot recognize the restriction sites and fragment the DNA at both the sites. Thus, DNA is not fragmented and remains in a linear form of 11 kb size. So, lane B represents the scenario that would result after digestion is performed on DNA in which mutations have altered both EcoRI site 1 and site 2. If 1000 bp DNA is inserted between the two restricted sites, then the size of DNA between the two restricted sites becomes 5 kb in size. Digestion of this DNA by EcoRI at site 1 and site 2 will result in 2 kb, 5 kb and 5 kb fragments. The 5 kb fragment DNA band intensity is more in lane E due to the two 5 kb DNA fragments that are generated after digestion of 1000 bp inserted DNA. So, lane E represents the scenario that would result after digestion is performed on DNA in which 1000 bp of DNA is inserted between the two restriction sites. If 500 bp DNA is inserted between the two restricted sites, then the size of DNA between the two restricted sites becomes 3.5 kb in size. Digestion of this DNA by EcoRI at site 1 and site 2 will result in 2 kb, 3.5 kb and 5 kb fragments. Lane D has a 3.5 kb fragment. . So, lane D represents the scenario that would result after digestion is performed on DNA in which 500 bp of DNA is deleted between the two restriction sites.

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