5. Consider the following plasmid (size 8000 bp), with restriction sites at the positions indicated: PstI...

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5. Consider the following plasmid (size 8000 bp), with restriction sites at the positions indicated: PstI Xhol 8 kb 2000 XhoI Probe EcoRI a) This plasmid is digested with the enzymes listed below. Indicate how many fragments will be generated in each case, and give the sizes of the fragments. Pstl Xhol Combination of PstI + Xhol + EcoRI (triple digest) b) Draw the banding pattern you would expect to observe if each of these digestions is loaded into a separate well of an agarose gel, and the fragments separated by electrophoresis. In the 2 first well you load a DNA marker (M) containing fragments with sizes of 1000 bp, 2000 bp, 4000 bp and 8000 bp. c) This gel is transferred to a membrane in a Southern blot experiment, and hybridised to a radioactively labelled 200 bp probe, which anneals to the plasmid DNA at the position indicated on the diagram above. Draw the autoradiographic profile you would expect to observe for the membrane. 7000 4200 5. Consider the following plasmid (size 8000 bp), with restriction sites at the positions indicated: PstI Xhol 8 kb 2000 XhoI Probe EcoRI a) This plasmid is digested with the enzymes listed below. Indicate how many fragments will be generated in each case, and give the sizes of the fragments. Pstl Xhol Combination of PstI + Xhol + EcoRI (triple digest) b) Draw the banding pattern you would expect to observe if each of these digestions is loaded into a separate well of an agarose gel, and the fragments separated by electrophoresis. In the 2 first well you load a DNA marker (M) containing fragments with sizes of 1000 bp, 2000 bp, 4000 bp and 8000 bp. c) This gel is transferred to a membrane in a Southern blot experiment, and hybridised to a radioactively labelled 200 bp probe, which anneals to the plasmid DNA at the position indicated on the diagram above. Draw the autoradiographic profile you would expect to observe for the membrane. 7000 4200

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Answer1 Plasmid vectors are vehicles for driving recombinant DNA into a host cell and are an important part of molecular cloning which is the process of building DNA molecules and introducing them int... View the full answer