View the step-by-step solution to:

A 9kb fragment of human DNA that has EcoRI sticky ends is cloned into the plasmid PBR007 at its single EcoRI restriction site.

A 9kb fragment of human DNA that has EcoRI sticky ends is cloned into the plasmid PBR007 at its single EcoRI restriction site. The resulting recombinant plasmid is digested with EcoRI and then, separately, with BamHI. In addition, a third digest is performed, in which EcoRI and BamHI are used together. The resulting fragments are run out on a gel. Below are the diagrams of the gel patterns from each digest:


EcoRI 10kb 9kb

BamHI 8kb 6kb 5kb

EcoRI + BamHI 7kb 5kb 4kb 2kb 1kb


A. How many kbs is the PBR007 plasmid without the insert?

B. How many EcoRI and BamHI sites are present in the newly formed recombinant plasmid?

C. Draw a restriction map of the recombinant plasmid. In your map, distinguish between the human DNA fragment and the plasmid. Indicate the restriction sites and the distances between the restriction sites.

D. If Southern blot analysis was performed on the double digest using the original 9kb insert as a probe, which bands would this probe hybridize to?

Recently Asked Questions

Why Join Course Hero?

Course Hero has all the homework and study help you need to succeed! We’ve got course-specific notes, study guides, and practice tests along with expert tutors.

-

Educational Resources
  • -

    Study Documents

    Find the best study resources around, tagged to your specific courses. Share your own to gain free Course Hero access.

    Browse Documents
  • -

    Question & Answers

    Get one-on-one homework help from our expert tutors—available online 24/7. Ask your own questions or browse existing Q&A threads. Satisfaction guaranteed!

    Ask a Question