14 - BIO 305 Problem set for lecture 14 Winter 2008 1 A...

Info icon This preview shows pages 1–3. Sign up to view the full content.

View Full Document Right Arrow Icon
BIO 305 Problem set for lecture 14 Winter 2008. 1. A circular plasmid is isolated and digested with Eco RI, Hind III, and Xho I, both separately and in different combinations. The fragments generated by these digestions were separated by size on an agarose gel. The results are: Eco RI HindIII Xho I 7 kb 9 kb 6 kb 2 kb 3 kb Eco RI/Hind III EcoRI/XhoI HindIII/Xho I all three 4 kb 5 kb 4 kb 3 kb 3 kb 2 kb 3 kb 2 kb 2 kb 1 kb 2 kb 1 kb Construct a map of this plasmid showing the locations and Eco RI, Hind III, and Xho I sites.
Image of page 1

Info icon This preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
2. You are trying to generate a genomic clone of the C. elegans lin-4 gene in the plasmid vector called pUC18. You have a 2 kb PCR product that contains both lin-4 and an unknown gene (gene X). Below is a map of pUC18 and the PCR product. (a) In order to clone just the lin-14 gene, which restriction enzyme(s) should you use to digest the DNA fragment and the vector? (b) You perform your restriction digests of both the vector and the PCR fragment. Next you inactivate your restriction enzymes and perform a ligation reaction by adding DNA ligase to a mix of all your restriction fragments. Assume that you use conditions so that the ligation products have at most one inserted fragment. How many different circular plasmids that carry the Amp R gene will be present in your ligation mix? (c) You use your ligation mixture to transform a strain of E. coli that lacks both the Amp R and the lacZ genes. You plate out your transformation on plates containing X-gal. Some
Image of page 2
Image of page 3
This is the end of the preview. Sign up to access the rest of the document.

{[ snackBarMessage ]}

What students are saying

  • Left Quote Icon

    As a current student on this bumpy collegiate pathway, I stumbled upon Course Hero, where I can find study resources for nearly all my courses, get online help from tutors 24/7, and even share my old projects, papers, and lecture notes with other students.

    Student Picture

    Kiran Temple University Fox School of Business ‘17, Course Hero Intern

  • Left Quote Icon

    I cannot even describe how much Course Hero helped me this summer. It’s truly become something I can always rely on and help me. In the end, I was not only able to survive summer classes, but I was able to thrive thanks to Course Hero.

    Student Picture

    Dana University of Pennsylvania ‘17, Course Hero Intern

  • Left Quote Icon

    The ability to access any university’s resources through Course Hero proved invaluable in my case. I was behind on Tulane coursework and actually used UCLA’s materials to help me move forward and get everything together on time.

    Student Picture

    Jill Tulane University ‘16, Course Hero Intern