Pglo Transformation Lab Report

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Transforming E. Coli Bacteria with the Gene GFP (green fluorescent protein) By Using pGLO As a Vector

Introduction:
Genetic transformation is the alteration of a cell by taking DNA from another organism and expressing those new characteristics from the organism of which it took the DNA from (Julie Carnagie and Leonard C. Bruno 2006). The cells that are able to take up DNA from other organisms are known as competent. This genetic transformation is used for many different things, from making medicines to DNA cloning (Brenda Wilmoth Lerner and K. Lee Lerner 2002). There are three ways that genetic transformations occurs. The first way is known as projectile bombardment. This means that cells are hit with projectiles that carry foreign DNA. The …show more content…
This gene must be turned on by using arabinose. Arabinose is a sugar that turns on the GFP biosynthesis pathway (Campbell 2009). The vector used in this experiment is pGLO. This is a plasmid that contains GFP. Is also contains an ampicillin resistance gene which allows us to find transformed cells. The plates that the pGLO is put onto contains ampicillin and only the cells that take up the plasmid will grow on the plate. This experiment shows that E. coli can be genetically transformation with the gene GFP and take on that gene to show its characteristics. The null hypothesis is that E. coli cannot be genetically transformed with GFP.
Materials and Methods:
(Weedman 2014)
To start off the experiment, two microcentrifuge tubes were labeled, one +pGLO and the other -pGLO. This pGLO is a vector used in the experiment to transfer the gene. A micropipetter with a fresh tip was used to transfer 250 µl of transformation solution into both of the labeled tubes. This solution containing calcium chloride was used because it increases the permeability of the plasma membrane. After putting the transformation solution into the tubes, then they are placed into an ice bath. Then +pGLO tube was

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process of transformation was green fluorescent protein. Two tubes were labeled “+pGlo” and “-pGlo” and 250 μl of CaCl was added and tubes were placed on ice. I used a sterile loop and extracted 3 large circular colonies of E. coli then swirled it into the +pGlo tube and repeated this process for –pGlo tube. Under UV light both tubes displayed no color difference. The tubes were placed on ice for 10 minutes then heat shocked at 42 ˚C for exactly 50 seconds in order for transformation to occur with…

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Purpose: This investigation is demonstrating the act of genetic transformation. We inserted a gene into the bacterium E. coli. Gene being a piece of DNA which provides the instructions for making a protein. The main chemical components used in this lab were calcium chloride, E. coli bacterium, a broth (made up of carbohydrates, amino acids, vitamins, nucleotides, and salts that allow bacteria to grow), pGLO plasmid (contains arabinose operon that turns on when the arabinose sugar is present, GFP…

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