Fill the chamber and cover the gel with buffer Make sure the wells of the gel

Fill the chamber and cover the gel with buffer make

This preview shows page 1 out of 1 page.

the electrophoresis chamber. Fill the chamber and cover the gel with buffer. Make sure the wells of the gel are near the black (-) electrode and not the red (+) electrode. Load 10 ul of each sample (L, P, E, H) into separate wells. Place the lid on the chamber and turn on the power to run the gel. When the electrophoresis run is at its completion, turn the power off and remove the gel from the tray into a staining tray. Stain the gel, rinse, and wash it to be able to visualize the DNA fragments. Results: (attached) Discussion: To determine the DNA fragment sizes, we first estimated the unknown DNA band sizes by comparing them to the known sizes on the gel. We then plotted the known distances vs. base pairs of HindIII (standard curve) to approximate the sizes of the unknown DNA. We could have made our DNA size estimation more accurate by running another DNA sample of known fragment size to create two standard curves as opposed to only one. We also could have run our gel longer. Both methods seemed to be somewhat accurate and inaccurate. Direct gel examination allowed us to estimate the sizes over the entire range, which was helpful for larger fragments that were beyond the curve’s capacity. The semilog graph allowed us to pinpoint almost the exact sizes of fragments, which would not be possible from just looking at the gel. Conclusion: Sources of error that may have changed the amount of fragments visible on the gel include running the gel for a longer period of time, using a different DNA stain, changing the concentration of the agarose for the gel, and changing the concentration of the buffer solution. Our results showed that there were five bands for Pstl, 3 bands for both EcoRI and HindIII and one band for the uncut DNA. There might have been more for the restriction enzymes, but since many of the bands migrated so closely together, they appeared as one band while others did not even appear. Pstl had smaller molecules that allowed it to run so far up the gel while the other two enzymes did not have as many.
Image of page 1

You've reached the end of your free preview.

Want to read the whole page?

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

Stuck? We have tutors online 24/7 who can help you get unstuck.
A+ icon
Ask Expert Tutors You can ask You can ask You can ask (will expire )
Answers in as fast as 15 minutes