Hi, the smallest recombination plasmid contains the lambda/HindII0.13kb insert.

Question

Hi,

the smallest recombination plasmid contains the lambda/HindII0.13kb insert. the size of this recombination is 2.82kb which is only slightly larger than the pUC18 vector (2.69kb) say you were given purified samples of both plasmids without being told which was which and ask to come up with a way to identified each plasmid. how would you accomplish this task? (clue: Two ways involving gels, the first a straight gel run of the plasmids but consider the composition of the agarose gel. The second way involves cutting the plasmids before using the same type of gel as for method one.)

Explanation / Answer

Running both the plasmids on agarose gel will give band of both plasmids nearer to the wells. This is so because plasmid DNA is supercoiled, so it will be retarded in the gel Although the retardation will be less for pUC 18 which is of smaller size, 2.69kb than for lambda which is 2.82kb

Cutting the plasmids with restriction endonucleases will accomplish the final task. Cut lambda with hind II0 and cut pUC 18 with HindIII. pUC will migrate farther in the gel.

Related Questions

Navigate

• Browse (All)
• Browse H
• Subjects

Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.