Graph of base pairs against the migrated distance of the fragments of DNA Conclusion: The fragments that moved more across the gel were the ones that had less base pairs because their mass is smaller and they can move across the gel more easily than the fragments that had more base pairs. So the first fragment in the gel is going to be the one with the most base pairs followed by the one with the second biggest number of base pairs.
The control was the one that moved less because it had the complete strand of the Lambda DNA and so is the one that had the most base pairs and it had more problem to move along the gel than the other stains that are only fragments of the Lambda DNA. Some of the fragments were “missing” because they were not stain with the dye or they are too small to be seen by the human eye. With the graph that was made using the data the average data of Hind III we can know how much would any fragment of the Lambda DNA of any size would move across the gel
Problems with the experiment: Some of the Lambda DNA was not stain or they were too small to be seen, this problem could be resolved by using a fluorescent gel that can stain more all the Lambda DNA fragments. Also some of the lines in the gel were too close to each other and it was had to identify them, this could be resolved by using a gel with a lower density than the one that we used.
Pipette the loading dye/DNA mixture into one of the wells, holding the tip above the well but under the buffer solution. Make a note of which DNA you have put into the well. Repeat the last two steps with each DNA digest and the ‘control’ tube with no restriction enzyme. Remember to use a new tip for each digest, to avoid cross-contamination. Fit one electrode at each end of the tank as shown in the illustration. Join the electrodes to batteries using wires with crocodile clips. Sufficient batteries should be used to give 18 volts.
Check that contact is made between the buffer solution and the electrodes. Leave the gel to ‘run’, undisturbed, for seven hours. Disconnect the batteries once the blue loading dye has reached the end of the gel. Rinse the crocodile clips in tap water and dry them thoroughly to prevent corrosion. Pour about 10 cm3 of staining solution onto the surface of the gel. Leave it for exactly 4 minutes.
Wash surplus stain from the gel surface with about 5 cm3 of 70% ethanol for a few seconds Pour away the alcohol, and then rinse the gel very carefully with cold tap water. Rinse with water 3 or 4 times to remove any excess stain. The remaining stain will gradually move down through the gel, staining the DNA as it does so. Faint bands should start to appear after 10 minutes. The best results will be seen if the gel is left to ‘develop’ overnight. (Put the tank in a plastic bag if you do this, to prevent the gel from drying out.).