Dna Synthesis Lab

When studying DNA it is required to break apart the genome at certain places in order to study certain sequences and see how they properly function and or possibly find the source of a mutation (1). Separating the strands require a certain protein to ensure that the rest of the DNA strand remains intact (2). Restriction enzymes are vital to this process because they are able to scan the DNA molecule very slowly to find the certain recognition site required to complete an experiment, which usually runs from four to eight nucleotides(1). Once the enzymes find the sequence they are able to stop and to bind to the DNA wrapping around the strands that contain the desired nucleotides separating it from the other sequences, also known as cutting the sequence (3). That

This experiment was completed by adding a PacI-linearized plasmid substrate to a master mix containing buffer and water, the same for each reaction. After, the SacI enzyme was added to each reaction with different characteristics. Thirdly, we took those incubated reactions and placed them into an agarose gel to test whether DNA fragments would separate using the process of electrophoresis. We hypothesized that the DNA will not be cut in reactions one, four, and seven due to there being no enzyme in reaction one. It will not cut in reaction seven because the enzyme was boiled which caused it to denature and lose its shape and once a protein loses its shape, it also loses its function not allowing it to cut the DNA. Reaction four was kept at a colder temperature which could result in the denaturing of the enzyme and the DNA not cutting properly. But we did believe that SacI would cut the DNA perfectly in reactions two, three, five, and six because the function enzyme was not affected by any of the alterations resulting in the cutting of