Analysis and Results
When the gels were finished incubating, I examined them on a light box. Using a centimeter ruler, I carefully measured each sample and the standards from the well to the end of the band and then recorded the distance. I then outlined the gel onto a clear transparency sheet, marking the wells, banding patterns, and standard patterns, for a permanent copy of results.
I analyzed the data by taking the average distance of the three runs per gel from each sample site. If the average distances equaled each other, then the LDH isoenzymes were the same. Thus the organisms had similar LDH enzyme composition between the two sites. If the migration distances were not equal, then LDH isoenzyme composition was different between the two sampling locations.
With the first gel, I did not run the Calf Serum standards with it because it had not come from the company yet. Instead, dye was used, to mark the migration. The results after the electrophoretic run showed that there was a difference between sites. There was a 2.44% migration difference in the samples. Meaning that the Thompson Valley extract traveled 2.24% further than the Bear's Grass extract did. In addition to the six bands I measured for the total distance, there were also some dark streaks occurring before the last band on the gel that the dye picked up. I sketched the outlines of those dark bands onto the transparency for my results, but I did not incorporate those measurements into this data because I mainly concerned myself with the maximum distance traveled by each extract because these bands were distinct and could be compared between the two sample sites. They also could indicate a difference in the sizes of the smallest isoenzymes between sites.
The second gel showed similar results to the first gel. This time, the Calf Serum was used as standards. The standards were in the first and last wells and after the electrophoresis, they measured equally to each other. Because the standards were similar on each side, this shows that the gel is even throughout and the LDH staining is working. I could only see the banding patterns from the first extract. They were all the same length. Banding patterns from the second extract could not be seen on the gel. Several factors could have effected the absence of the bands, which include: they ran out of the gel, the dye didn't pick up the bands, or there was not enough protein to be seen. Due to the fact that there were no bands in the second extract on this electrophoretic run, the percentage of migration difference is not applicable to this gel. The similarity between gel one and gel two were the dark streaking patterns before the band. The dark streaking patterns occurred in all six of the wells containing extract. The streaks are noted on the transparency.
Gel number three had banding patterns that were different lengths between locations. The Bear's Grass migration distances were longer than the Thompson Valley migration distances. The percent difference in migration distances was 24.44%.
The fourth gel also had banding patterns that were different lengths between locations. Bear's Grass had migration distances that were longer than the Thompson Valley migration distances. The percent difference in migration distances was 12.50%.
I ran a total of four gels with three samples per location for comparison. Therefore the organisms from each location were analyzed for LDH isoenzymes a total of 12 times. The following graph represents the distance of migration from each well for the four gels.
