Posted: May 22nd, 2023
Lane 4, which contained HindIII, acted as the guide for the sizes of DNA cut. HindIII is normally consistent and acts as a reliable guide when comparing results obtained from different procedures. It is noted that in the lane, it is only seven bands that are observed clearly as the other one is too small and most probably run off the gel.4[228]Â Because of the inverse log scale of the cuts and the percentage of agarose used, it is hard to separate the two large cuts. As a result of the consistency achieved from HindIII lane, it is easy to determine where the cuts by the other enzymes were made.
The first lane contains uncut lambda DNA. It is easy to determine this since only one band is observed in the lane. The lane contained no enzyme to act on the DNA. In the case of lane 2, BamHI was expected to have six bands, but only five are shown in the results. The results may be due to the fact that two pairs of the expected cuts, 5505bps and 5626bps, as well as 6527bps and 6770bps, are so close to each other. The proximity of the cuts to each other makes them appear as one. The results are similar to those observed in the mystery enzyme. Therefore, a conclusion is made that the enzyme is BamHI.
For lane 2 that contains E Cori, the expected number of fragments is 6, but only 5 are showing.3[1049] The two bands located at 5643bps and 5804bps are viewed as one big and bright band located just below the 6557bps point of HindIII. The last lane contains the experimental enzyme used in the experiment. Three bands are observed, the first at roughly between 23130bps and 48502bps. The second one is located just below 20000bps and above 9416bps. The last band is observed at just below 6557bps at around 5500bps. The enzyme matches the action of BSa1, which produces fragments at 31291bps, 11418bps, and 5793bps.4[241]
There are two reasons as to why some bands are darker than others. The first reason is that the larger fragments contained more ethidium bromide than the smaller ones because there were more places for it to be inserted.3[1053]Â As a result, the larger fragments are brighter compared to the smaller ones. The second reason is that the dark regions were produced by the loading dye, which bleached out some of the florescence of the ethidium bromide.
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