The result from the transformation process showed no colony growth on the agar plate, which means no plasmid DNA got inserted into the vector (Table 1). Thus, the transformation was considered unsuccessful. With that being said, the gene was not transferred among the bacterial cells. The reason could have been that the agar plate had a strong concentration of antimicrobial drug, so the bacteria may have been unable to grow under such condition. Another reason could be that the cell did not get enough heat shock, or not enough pre-incubation time with CaCl2, which means the bacterial cell wall was not permeable enough for the DNA to come inside and interact with the bacterial DNA.
The DNA gel electrophoresis showed that there were no fragments …show more content…
However, the concentration of both PCR samples were measured using the nanodrop machine, and the result showed that the concentration for PCR sample A and B were 651 and 233 ng/μL, respectively. It proved that there was DNA cloned in the samples. Another DNA gel was redone for the PCR sample (Fig 2). The second gel showed that there was a light band of PCR sample B at around 300 base pairs. It matched with the expected data for PCR; however, PCR sample A did not show any band. It was concluded that the DNA gel electrophoresis was not successful. Some error occurred during the PCR process, and it could be improper mixing of the reagents, since the amount of each reagent was small. Improper mixing would lead to a missing reagent, and therefore PCR would not turn out right. Another error could be that the concentration of DNA was low, so there was not enough DNA to migrate through the …show more content…
The gel indicated that in flow of fraction 1, the band had migrated all the way down the ladder. The multiple bands in the result means it contained many impurities. For fraction 2 with TEA wash, the bands migrated the same length as the first fraction which implies that it also contained impurities. For fraction 3, 5 mM of imidazole was eluted with the solution and it showed a light band at 38 kDa. It means there was presence of proteins in this fraction. For fraction 4, a higher concentration of imidazole was used to wash the solution, the gel showed the band migrated and stopped at 38 kDa. For fraction 5, highest concentration of imidazole was used and the migration stopped at 38 kDa as well. It represented the presence of purified protein in the fraction by showing only one band at the site, which indicated the protein was purified. The expected SpHTS protein was around 40 kDa. In the gel, the purified protein was at 38 kDa, which showed the protein was purified (Fig
The DNA gel electrophoresis showed that there were no fragments …show more content…
However, the concentration of both PCR samples were measured using the nanodrop machine, and the result showed that the concentration for PCR sample A and B were 651 and 233 ng/μL, respectively. It proved that there was DNA cloned in the samples. Another DNA gel was redone for the PCR sample (Fig 2). The second gel showed that there was a light band of PCR sample B at around 300 base pairs. It matched with the expected data for PCR; however, PCR sample A did not show any band. It was concluded that the DNA gel electrophoresis was not successful. Some error occurred during the PCR process, and it could be improper mixing of the reagents, since the amount of each reagent was small. Improper mixing would lead to a missing reagent, and therefore PCR would not turn out right. Another error could be that the concentration of DNA was low, so there was not enough DNA to migrate through the …show more content…
The gel indicated that in flow of fraction 1, the band had migrated all the way down the ladder. The multiple bands in the result means it contained many impurities. For fraction 2 with TEA wash, the bands migrated the same length as the first fraction which implies that it also contained impurities. For fraction 3, 5 mM of imidazole was eluted with the solution and it showed a light band at 38 kDa. It means there was presence of proteins in this fraction. For fraction 4, a higher concentration of imidazole was used to wash the solution, the gel showed the band migrated and stopped at 38 kDa. For fraction 5, highest concentration of imidazole was used and the migration stopped at 38 kDa as well. It represented the presence of purified protein in the fraction by showing only one band at the site, which indicated the protein was purified. The expected SpHTS protein was around 40 kDa. In the gel, the purified protein was at 38 kDa, which showed the protein was purified (Fig