Introduction:
Genetic transformation refers to the procedure in which a portion of foreign genetic material is implanted into a cell and becomes evident through change in appearance or function. This process may appear naturally, or by methods such as projectile bombardment, electroporation, or heat shock. Projectile bombardment occurs when cells in a culture are blasted by pellets that are covered with the DNA they are hoping to insert into the cell. It works as a type of gene gun. Electroporation takes place when cells exist in a liquid medium that contains the foreign DNA and are shocked by a series of electrical pulses, forcing the plasma membrane to become more permeable and take up the DNA. In heat shock, the method we used in this report, …show more content…
In the case of bacteria, a sugary substance known as arabinose must be present in order for the GFP to be physically active and visible. The arabinose C gene also present with the GFP recognizes and binds to arabinose so that they gene may produce the glow. A common bacteria used in this process is Escherichia coli, better known as E. coli. It is a small, single-celled bacterium with a rapid reproduction rate, making it ideal to use in transformations since the effects will be seen much faster than with other hosts. When the bacterium receives the genes from the plasmids, it provides better traits for adaptation and survival, but it is only successful if all substances needed are in the mixture as well (Masterman 2010). My hypothesis is that using the heat shock method described above, the plasmid pGLO will be used as a vector to transfer green fluorescent protein, arabinose C, and ampicillin resistance from the pGLO to the E. coli bacteria and will present itself by glowing in the presence of arabinose and surviving on media that contains ampicillin. The significance of this experiment is to guide students through using the scientific method to complete a laboratory exercise, specifically determining whether or not the properties of pGLO will be active in …show more content…
Both tubes were labeled, one being positive pGLO, and the other negative pGLO. Then, using a long sterile loop, each group picked up a single colony of E. coli that was then mixed throughout the positive tube and returned back to the ice. The same was then done for the negative pGLO tube. With a new sterile loop, each lab group obtained a sample of pGLO plasmid DNA and submerged it into only the positive tube and placed it back into the ice. Each group then received four Luria Bertani broth nutrient agar plates and labeled them +pGLO LB/amp, +pGLO LB/amp/ara, -pGLO LB/amp, and –pGLO LB, the amp standing for ampicillin and the ara for arabinose. After both the tubes had been chilled in the ice for ten minutes they were placed on small floatation devices and placed directly into a tub of 42 degree Celsius water for no more than 50 seconds, simulating the heat shock treatment. Immediately after removing them from the water, the tubes were back in the ice filled beaker for another two minutes. When the two minutes was up the tubes were taken out of the ice and each had 250 μl of nutrient broth mixed in them and left to sit at room temperature for ten minutes. After sitting for all
Genetic transformation refers to the procedure in which a portion of foreign genetic material is implanted into a cell and becomes evident through change in appearance or function. This process may appear naturally, or by methods such as projectile bombardment, electroporation, or heat shock. Projectile bombardment occurs when cells in a culture are blasted by pellets that are covered with the DNA they are hoping to insert into the cell. It works as a type of gene gun. Electroporation takes place when cells exist in a liquid medium that contains the foreign DNA and are shocked by a series of electrical pulses, forcing the plasma membrane to become more permeable and take up the DNA. In heat shock, the method we used in this report, …show more content…
In the case of bacteria, a sugary substance known as arabinose must be present in order for the GFP to be physically active and visible. The arabinose C gene also present with the GFP recognizes and binds to arabinose so that they gene may produce the glow. A common bacteria used in this process is Escherichia coli, better known as E. coli. It is a small, single-celled bacterium with a rapid reproduction rate, making it ideal to use in transformations since the effects will be seen much faster than with other hosts. When the bacterium receives the genes from the plasmids, it provides better traits for adaptation and survival, but it is only successful if all substances needed are in the mixture as well (Masterman 2010). My hypothesis is that using the heat shock method described above, the plasmid pGLO will be used as a vector to transfer green fluorescent protein, arabinose C, and ampicillin resistance from the pGLO to the E. coli bacteria and will present itself by glowing in the presence of arabinose and surviving on media that contains ampicillin. The significance of this experiment is to guide students through using the scientific method to complete a laboratory exercise, specifically determining whether or not the properties of pGLO will be active in …show more content…
Both tubes were labeled, one being positive pGLO, and the other negative pGLO. Then, using a long sterile loop, each group picked up a single colony of E. coli that was then mixed throughout the positive tube and returned back to the ice. The same was then done for the negative pGLO tube. With a new sterile loop, each lab group obtained a sample of pGLO plasmid DNA and submerged it into only the positive tube and placed it back into the ice. Each group then received four Luria Bertani broth nutrient agar plates and labeled them +pGLO LB/amp, +pGLO LB/amp/ara, -pGLO LB/amp, and –pGLO LB, the amp standing for ampicillin and the ara for arabinose. After both the tubes had been chilled in the ice for ten minutes they were placed on small floatation devices and placed directly into a tub of 42 degree Celsius water for no more than 50 seconds, simulating the heat shock treatment. Immediately after removing them from the water, the tubes were back in the ice filled beaker for another two minutes. When the two minutes was up the tubes were taken out of the ice and each had 250 μl of nutrient broth mixed in them and left to sit at room temperature for ten minutes. After sitting for all