Plasmid Practice Problems Key

Then a polylinker was designed to be first inserted

Info icon This preview shows pages 1–4. Sign up to view the full content.

View Full Document Right Arrow Icon
Then a polylinker was designed to be first inserted into the SwaI site, after which the 5’ arm will be subsequently cloned into the ploxPFlpneo plasmid utilizing one of the restriction sites in the “SwaI polylinker.” The SwaI polylinker sequence is shown below in capital letters: agacgcATTTAAATTCGAATTCCTCAGCAGATCTGGGCCCATTTAAATagacgc In the Linker: BbvC I = CC*TCAGC Bgl II = A*GATCT BstB I = TT*CGAA EcoR I = G*AATTC Apa I = GGGCC*C Swa I = ATTT*AAAT Outside the Linker: Swa I = ATTT*AAAT BamH I = G*GATCC Apa I = GGGCC*C Xho I = C*TCGAG * denotes where the restriction site is cut by the restriction enzyme.
Image of page 1

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

View Full Document Right Arrow Icon
ploxPFlpneo plasmid schematic
Image of page 2
Questions: 1. What are some potential reasons as to why the 5’ arm was so difficult to clone into the SwaI restriction site (before the SwaI polylinker was introduced)? SwaI is a “blunt cut” restriction enzyme. Blunt cut restriction enzymes are less efficient than “sticky end” restriction enzymes. As such, larger DNA fragments are difficult to ligate in between SwaI and more ligase is required. 2. What is the most likely reason why the 5’ arm must be cloned into this plasmid before the 3’ arm? There is a SwaI restriction site within the 3’ arm; conversely, there is no XhoI site in the 5’ arm.
Image of page 3

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

View Full Document Right Arrow Icon
Image of page 4
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