Molecular Biology Lab Report Essay

1660 Words Apr 5th, 2011 7 Pages
Molecular Biology Lab Report
Mapping DNA using Restriction Enzymes Ava II and Pvu II to cut Bacterial DNA
The objective of this project is to map bacterial DNA, which is derived from E. coli, using restriction endonucleases with gel electrophoresis. The DNA fragments, after cutting has occurred, are separated using agarose gel electrophoresis. The DNA fragments are placed in the gel, and an electric current is run through the matrix of the gel-like agarose. Migration of the fragments across the gel is based on the size and charge of the fragment. After the fragments have been run in the gel, they are stained with methylene blue and viewed on a light box. This allows the DNA bands to be viewed on the gel.
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e. The gel was poured slowly into the gel plate. The solution covered half of the height of the comb’s teeth. Bubble formation was prevented. In case of formation of bubbles, a sharp object could be used to carefully burst them. f. The gel was allowed to solidify. It became opaque when solidified. g. After the gel had solidified, water was added sparingly around the comb and the comb was removed slowly and evenly. Wells were created by the removal of the combs.

4. Load the gel and do electrophoresis of samples a. The gels were placed in the running chamber with the wells closest to the negative (black) electrode, and the gel was completely covered with 1X TBE. The wells pointed outwards so as to enable better viewing. b. Three additional samples were collected from the instructor. These samples were labeled 1 (the loading dye), 2 (molecular weight markers), and 6 (uncut pUC 19 DNA). c. The six microtubes were assembled with their respective solutions and were arranged in order in a microtube rack. d. 2 µL of loading dye were added to each of the six tubes. e. 2 µL of pUC 19 DNA that had been uncut was added to tube 6. f. Each of the six samples was loaded carefully into their corresponding wells. g. The positive and negative electrodes were carefully attached to their respective terminals on the power supply and on the gel box. h. The power was set to about 100 volts. i.

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