October 29, 2014
Biology 1 Cellular Processes Lab
Materials and Methods
Part I – Restriction Enzyme Digestion
To begin this experiment, the DNA molecules must be cut into smaller fragments with distinct enzymes called Restriction Enzymes through a process called Restriction Enzyme Digestion. Four microtest tubes were labeled 1 through 4 and added 10 µl of Enzyme Reaction Buffer to each of the four reaction tubes using a micropipette. DNA, and Enzyme 1 and 2, were then added to the reaction tubes using a new micropipette tip for each transfer of DNA and enzyme (refer to figure 6.1). The solutions were covered and mixed gently by tapping the reaction tubes on the table which then allowed for the contents to assemble at the bottom of the reaction tubes, followed by incubating the reaction tubes in a 37°C waterbath for 45 minutes. Finally, after the incubation was complete, 5 µl of 10x gel loading solution was added to each reaction tube to stop the reaction. Once again, the reaction tubes were covered and tapped gently to mix the solution and stored in the refrigerator for gel electrophoresis.
Table 6.1 Summary of Restriction Enzyme Digestion …show more content…
Gel electrophoresis is used in day-to-day activity in various ways such as maternity/paternity testing, and genotyping, testing for heritable diseases. DNA typing is also used in Identifying human remains. Scientists have used DNA profiling to verify or refute that the person in the grave was or was not, the person who was believed to be there. DNA typing can likewise be manipulated to identify food, for example, fish supplied in sushi meat was in actuality from dolphins and