Pglo Lab Report

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In transforming the genetic makeup of bacteria to make E.coli glow takes a lot of steps. For this experiment you will need, 1 E.coli starter plate, 4 poured agar plates (1 LB, 2 LB/amp, and 1 LB/amp/ara), transformation solution, LB nutrient broth, inoculation loops, pipets, foam microcentrifuge tube holder/float, container of crushed ice, marking pen, copy of guide, microcentrifuge tubes, rehydrated pGLO plasmid, 42*C water bath and thermometer, UV light, 37*C incubator, 2-20 ul adjustable volume micropipets, and 2-20 ul micropipet tips.
First, you need to label 2 micro test tubes +pGLO and -pGLO. This needs to also have your groups initials or names if you are doing a class experiment. If you are doing a lab experiment for work, you should put …show more content…
Second, you need to add 250 ul of transformation solution (CaCl 2) into each tube with a sterile loop. If the loop is plastic, change the loop each time you need one. If the loop is metal, you don’t need to use a new one each time, but you need to make the loop sterile each time by burning the loop until it turns bright red. This is what will increase the ability to transform the DNA. Third, place the tubes in the tube rack and then place the tube rack in a bowl of ice cubes. With this step, leave the tubes open. For this experiment to work the ices has to be crushed not cubed. Fourth, using a new sterile loop or a freshly sterilized loop, place 2-4 large colonies of bacteria from your starter plate. It is very important to take only a single colony not a swab of bacteria. Pick up the +pGLO tube and place the bacteria in the tube. Then spin the tube in between your index finger and your thumb until it is evenly spread out and there are no floating chunks. Place the tube back in the rack and back on

Pglo Lab Report

centrifuge tubes were individually labeled –pGLO and +pGLO and then placed into a foam tube rack. Each tube was then opened and given approximately 250 µL of the transformation solution (calcium chloride), through the use of a micropipetter (with a fresh tip for each). Subsequently both tubes were closed and placed directly in a tub of ice. A single colony of bacteria was then picked up from the starter plate by a sterile loop and placed in the +pGLO tube. The loop was twirled in the transformation…

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+ Pglo Lab Report

coli colonies you initially observed? Explain your predictions. Plates 3 and 4 should contain bacteria that are most like the original non-transformed E. coli colonies. This is because these plates serve as control plates. They do not contain the pGLO plasmid that causes genetic mutations or transformations. Plate 3 should not contain any bacteria because it has ampicillin, which prevents bacteria growth, but plate 4 should contain bacteria because it does not contain ampicillin. 2. If there…

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Pglo Lab Report

Purpose The overall purpose of the pGLO lab was to transform E. Coli bacteria by adding materials such as luria broth, ampicillin, plasmid, and transformation solution. Transformation is a process in which one strain of bacteria is changed by a gene or multiple genes from another strain of bacteria. Procedures The first thing done for this lab was prepping the plates. Students labelled the plates, and then teachers put in agar agar in each plate, adding either pGLO plasmid, ampicillin, or arabinose…

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Pglo Transformation Lab Report Essay

Transformation Lab Report Introduction Transformation is the transfers of virulence from one cell to another, through the transferring of genetic material. It was originally postulated in 1928 through the works of Federick Griffith, a British microbiologist. Griffith observed that the mutant form, non-virulent form, of the bacteria Streptococcus Pnumoniae could be transformed into the normal, virulent form, when injected into mice along with heat killed normal forms. He concluded that somehow…

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+ Pglo Transformation Lab Report

The protein used in 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…

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Pglo Transformation Lab Report

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…

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Pglo Transformation Lab Report

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 that glows green in fluorescent light, bla that provides ampicillin resistance,…

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Pglo Transformation Essay

Connor Lauffenburger 3/17/13 pGlo Transformation Lab Report I Introduction The purpose of this experiment was to show the genetic transformation of E. coli bacteria with a plasmid that codes for Green Fluorescent Protein (GFP) and contains a gene regulatory system that confers ampicillin resistance. A plasmid is a genetic structure in a cell that can replicate independently of chromosomes. In this lab, the Green Fluorescent Protein, which is typically found in the bioluminescent jellyfish Aequorea…

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Lab Report Guidelines Essay

GUIDELINES FOR LAB REPORT Concise and Informative Title that Reflects The Findings of the Experiment [Arial font; 12 pt, bold, centered) A. Author and B. Author (MARB 301, Section 40x) [Arial font; 10 pt, bold, centered] Texas A&M University at Galveston, Genetics Lab (MARB 301), 200 Seawolf Parkway, Classroom Laboratory Building, Galveston, TX 77553 Summary Provide a brief summary of your findings in general terms that will entice me to read on  [Font: Arial, 10 pt*, left aligned] Up to 100 words…

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Essay about Pglo Transformation Lab Final 1

pGLO Transformation Lab In this lab you will perform a procedure known as genetic transformation. This term means change caused by genes; it involves inserting a gene into an organism in order to change one or more characteristics in that organism. We will use E. coli K-12 as our host organism; ie the organism that will receive the gene. The gene we will transfer to E. coli K-12 codes for green fluorescent protein (GFP). The source of this gene is the bioluminescent jellyfish Aequorea Victoria…

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