Using The Uv Box And Its Effect On The Body Essay

Using The Uv Box And Its Effect On The Body Essay

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The picture taken using the UV box is shown in the figure below. We could see that the restriction digests were successful as it is clear and easy to visualize the different bands and fragments separated, however, the separation of the ladder bands is not very clear. Thus it would be hard to rely on these results to determine the sizes of the fragments. The blurry ladder bands could have resulted from the amount of time the gel was running for, the voltage or even the way it was loaed into the well. Along with the ladder bands, the DNA bands are not very bright although the separation of the fragments is clear (which is the most important part). This could probably have been related to the running buffer used.

Figure 1: photograph of the gel after electrophoresis

We can also see that the DNA bands have some bright dots in the center. Those are a result of the way the loading was performed. When the tip is very close or touches the bottom of the well, the DNA in the mixture is then concentrated in the one spot at the center when being pipetted out. This does not effect by any means the running of the bands but is more of a visual manifestation. Given all what have been said, we are not going to rely on this gel generate our data mainly because the ladder bands are not very clear. Therefore, an alternative gel is going to be used as to generate the standard curve and to determine the sizes of the fragments observed in the gel and to eventually attempt the different plasmid maps for giving the data resulting from each reaction. The figure below represents the alternative gel used, showing all the distances travelled by the samples loaded which are the distances measured from the center of each band to the bottom of the wells.


... middle of paper ... plasmid maps to represent each of the restriction sites for each of the enzymes used. The three plasmid maps with a single enzyme used are shown below.
HindIII PvuII BglI

The three plasmid maps with double restriction enzymes used are as follows:
HindIII + BglI PvuII + BglI PvuII + HindIII

Combining all plasmid maps with single and double enzymes, we can obtain a more general map with all restriction enzyme involved and representing all the recognition sites.
HindIII + BglI + PvuII

In the next section, we will analyze the results obtained from measuring the distances traveled and explain how the different molecular weights of all bands allowed us to figure out how many fragments are generated in each reaction and to construct the different plasmid maps. Potential experimental problems or deviations will also be discussed.

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