Answers are provided in the Teachers Answer Guide. pGLO™ Bacterial Transformation Kit Catalog #166-0003EDU explorer.bio-rad.com For technical support call your local Bio-Rad office, or in the U.S., call 1-800-424-6723 pGLO araC GFP bla ori See individual components for storage temperature. Transformation is the introd… Without bacteria, we would not be able to digest food or produce some of our favorite foods such as yogurt and cheese. The UV light is necessary to cause the GFP protein within the bacteria to fluoresce. Spell. The plasmid must express a gene for ampicillin resistance (the protein product of the bla gene codes for beta-lactamase, the protein that breaks down ampicillin). State the purpose of each component of the experiment, inhibits cell growth by interfering with cell wall synthesis, regulate Green Fluorescent Protein (GFP) expression in transformed cells, transforms bacteria that is used to make a cell competent, increases, possesses the ability to absorb blue light and in response show green, Substance that allows bacteria to recover before being plated. pKwi Bacterial Transformation Q1 Answers to Questions Answers to Questions: Teacher’s Edition PreLab Activity: Student Learning Outcome & Pre-Lab Predictions For Laboratory Activity Student Learning Outcomes – at the end of this laboratory, students will be able to: 1. HUMA KHAN 4/27/20 LAB 10 QUESTION ANSWERS Lab 10: Bacterial Transformation. Safety is another important consideration in choosing an experimental organism. an organism to change the organisms trait. These bacteria were removed from the starter plate, did not have any plasmid added to them, and were replated on an LB plate. 1285 Bacterial Transformation Virtual Lab - A.P. By introducing a new gene into an organism via a vector (DNA molecule used as a vehicle to artificially carry foreign genetic material into another cell) (plasmid, virus, etc.) You cannot tell if the bacteria are ampicillin resistant just by looking at them. Explain the process of bacterial transformation. Good examples of highly regulatable genes are the enzymes which break down carbohydrate food sources. This process creates a uniform electrical field that allows motion of particles of various sizes towards a positively charged end. pGlo plasmids, when taken up by a bacteria, will code for. LabBench Activity Analysis of Results I. What is meant by the control plate(s)? Created by. The colonies on the LB/amp/ara plate fluoresce green under UV light, and the transformed colonies can grow on ampicillin resistance. The growth of E. coli on the two plates could be compared. Gravity. A bacterial colony is a large group or cluster of bacterial cells that originated from a single, clonal cell. Course. I just need help with formulating the hypotheses. Although the E. coli strain used in these experiments has been rendered non-pathogenic, it is important to teach the students good sterile technique and safe disposal of bacteria. Instead of having students answer the "cookbook" pre-lab and post-lab questions where they can find the answers online, I have them write a formal lab report.The resourc. Yes. Bacteria which resemble the non-transformed will be found on the LB/(-) pGLO plate. How would you change the bacteria's environment to best tell if they are ampicillin resistant? PLAY. What are 3 real-world links for the study/use of genetic GFP? To get this information, which would be a better candidate for your investigation, an organism in which each new generation develops and reproduces quickly, or one which does this more slowly? Colony size is similar both before and after transformation. This is why you remain in the best website to look the incredible book to have. fatpanda80. Scientists often want to know if the genetically transformed organism can pass its new traits on to its offspring and future generations. 1 Ms. Strachan/AP Biology Bacterial Transformation (Virtual Lab) Table 1. Arizona State University. Roanne. Bacterial Transformation Lab Answers. The bacteria on the (+) pGLO LB/amp plate and the (-) pGLO LB plates should be whitish. If the genetically transformed cells have acquired the ability to live in the presence of the antibiotic ampicillin, then what can be inferred about the other genes on the plasmid that were involved in your transformation procedure? mocha_dog529. Only transformed cells can grow on agar with ampicillin. No. PLAY. Describe your reasoning. Bacterial Transformation Lab: pGLO. Providing publishers with the highest quality, most reliable and cost effective editorial and composition services for 50 years. This plate does not contain arabinose which is needed to induce expression of the GFP gene and generate green fluorescent colonies. 2. Learning Goals: Transform bacteria using the pGLO lab procedures Big Ideas: To develop newly acquired skills in the laboratory. Antibiotics usually kill bacteria (are bacteriocidic) or inhibit their growth (bacteriostatic). Woodstock High School, Woodstock • SCIENCE 1035B, Truckee Meadows Community College • BIOLOGY 190, Isaac Perez Quintero - Transformation lab.pdf, University of California, Davis • BIOLOGY 2A. Flashcards. This plate contains arabinose which induces expression of the GFP gene and generates green fluorescent colonies. The organism should not be able to infect plants or animals. Relate the use of bacterial transformation in biotechnology. The best test would be to take some of the bacteria growing on the LB plate and streak them on an LB/amp plate. 24 2. This Site Might Help You. The organism should not produce any toxins or compounds which could make people sick. This exogenous DNA can be recombinant DNA molecules that have been constructed in vitro, as well as natural DNA molecules. In this investigation, students will first acquire the tools to transform E. coli bacteria to express new genetic information Page 3/5. In the Transformation Lab designed by the Carolina Biological Supply Co., we took extracted DNA and inserted them into E. Coli bacterial cells through the transformation process (Carolina Biological Supply Co. 2014). What happens to an unopened soft drink when it freezes? State the purpose of each component of the experiment Components: 1. Since only some of the cells exposed to the amp R plasmids will actually take them in, only some cells will be transformed. The (-) pGLO/LB control plate can be compared to any of the LB/amp plates to show that plasmid uptake is required for the growth in the presence of ampicillin. Attleboro, MA 02703 (508) 222-5150 ext. a green fluorescence protein GFP. Please sign in or register to post comments. STUDY. When lab is complete, collect all p… What do you think each of the two environmental factors you listed above is doing to cause the genetically transformed bacteria to turn green? Equal amounts of cells could be plated on two different LB nutrient agar plates, one which contains just LB nutrient agar and one which contains LB nutrient agar ampicillin. As this biology bacterial transformation lab answer key, it ends taking place brute one of the favored books biology bacterial transformation lab answer key collections that we have. Title: pGLO Transformation Lab Introduction: Genetic transformation is a change caused by genes, involving the insertion of a gene into. Thus you will see only individual colonies on the plate. Cells which were not treated with DNA (-pGLO) should not be expressing the ampicillin resistance gene and will not grow on the LB/amp plates. The plasmid sample (LB/amp +pGLO) did not fluoresce. Classzone Bacterial Transfomation Virtual Lab Answer Key What purpose does a control serve? 2. The ara control region regulates GFP expression by the addition of arabinose, so the GFP gene can be turned on and off by including or omitting arabinose from the culture medium. To genetically transform an entire organism, you must insert the new gene(s) into every cell in the organism. Genetic transformation occurs when a cell takes up and expresses a new piece or foreign ____________, often a circular plasmid. Terms in this set (12) What is bacterial transformation? Success Criteria: I can successfully transform the bacteria and investigate the heat factor in bacterial transformation. Describe the evidence that indicates whether your attempt at performing a genetic transformation was successful or not successful. Bacteria are a whitish color. There can be 10 to 200 copies of the same plasmid within a cell. Before undergoing the transformation lab, confirmation that the substance being added to the bacterium is DNA must be acquired. Of the E. coli traits you originally noted, which seem now to be significantly different after performing the transformation procedure? Do you observe some E. coli growing on the LB plates which do not contain ampicillin/arabinose? The bacteria need to be ruptured in order to release the GFP/plasmid DNA, which can then be purified using column chromatography. Flashcards. To ensure a pure culture, we must start with a single bacterium. The sugar arabinose turns on expression of the GFP gene by binding to a regulatory protein, araC, which sits on the PBAD promoter. Learn. Bacterial Transformation LAB Analyzing Results. What two factors must be present in the bacteria's environment for you to see the green color? Fast production of offspring or new progeny will allow you to quickly assess if the new trait has been passed on. Which of the two possible sources of the fluorescence can now be eliminated? Addresses AP® Biology Big Ideas 1 and 3 and Essential Knowledge 1.A.1, 1.A.2, 3.A.1, 3.C.1, and 3.C.2. The bacteria we might generally call "bad" for us include those responsible for causing illnesses like food poisoning. What is the total volume of reagent in mL? In the space below, list these non-transformed traits and how you arrived at this analysis for each trait listed. From your results, can you tell if these bacteria are ampicillin resistant by looking at them on the LB plate? Bacterial Transformation Lab Answers bacterial plasmid-based genetic transformation, enables students to manipulate genetic information in a laboratory setting to understand more fully how DNA operates. Bacteria are small, single-celled organisms which reproduce quickly and easily. Thus, there should be few, if any, bacterial colonies present on the ampicillin plate. Why? Explain your prediction. On which of the plates would you expect to find bacteria most like the original untransformed E. coli colonies you initially observed? On which of the plates would you expect to find bacteria most like the original nontransformed. When a bacterial cell freezes, the volume of cytoplasm expands. There are several techniques available to achieve this. List all observable traits or characteristics that can be described. Test. This is a lab report assignment to accompany the pGLO Bacterial Transformation Lab done in many AP Bio and Biotechnology classes. pGlo plasmids, when taken up by a bacteria, will code for. © Copyright, Cold Spring Harbor Laboratory.All rights reserved. 1. Related documents. Each colony can be seen by the naked eye, while a single bacterium requires a micro-scope for observation. E. coli colonies you initially observed? They also can explore answers to questions about plasmids and transformation that might have been raised during the initial investigation. The color of colonies, number of colonies, distribution of colonies on the plate. 2017/2018. 5 years ago. Duplication of any part of this document is permitted for classroom use only. Bacterial Transformation Lab? 1. From the results that you obtained, how could you prove that these changes that occurred were due to the procedure that you performed? The multiplication of a single bacterium on agar plates appears as a colony. Thus, the plasmid must confer resistance to ampicillin. The bacteria that did not receive the plasmid are growing on a plain LB plate. Match. Moreover, the colonies on the LB/amp/ara plate should fluoresce green. when a host organism takes in foreign DNA and expresses the foreign gene. If a white colony was streaked onto an LB/amp/ara plate, the resulting colonies would be green. What is the function of the “-DNA” tube. If there is no ampicillin in the agar, E. coli will cover the plate with so many cells it is called a "lawn" of cells. This document contains the questions for this lab that, are also included in the Bacterial Transformation Lab PDF. An unsuccessful experiment will show an absence of colonies on the (+) pGLO LB/amp and (+) pGLO LB/amp/ara plates. Which organism is better suited for total genetic transformation-one composed of many cells, or one composed of a single cell? 1. This usually occurs with plasmids, small circular molecules of DNA. If ampicillin has no effect, there should be approximately equal numbers of colonies on both plates. Subjects: Science, Biology, General Science. The LB/amp (-) pGLO and the LB/amp (+) pGLO plates should be directly compared. (Hint: one factor is in the plate and the other factor is in how you look at the bacteria). In the lab, we incorporated the plasmids into the bacteria and put them into a medium with an antibiotic. Genetically transformed cells have taken up the pGLO plasmid which expresses the ampicillin resistance gene—these cells can survive on the plates which contain ampicillin. The source of fluorescence is probably from some protein that the plasmid encodes from the addition of arabinose, namely GFP. Explain your answer. In this experiment, bacteria will be transformed with a gene that codes for Green Fluorescent Protein (GFP). Before any change in the phenotype of an organism can be detected, a thorough examination of its usual (pre-transformation) phenotype must be made. The (-) pGLO LB/amp plate shows that the starter culture does not grow on the LB/amp plate. Look again at your four plates. Can you predict what would happen if you took one of the green colonies from the LB/amp/ara plate and streaked it onto an LB/amp plate? This preview shows page 1 - 2 out of 3 pages. Thus, they are virtually identical to the non-transformed starter colonies. What was the purpose of rupturing or lysing the bacteria? The questions go in the same order that they are asked in the PDF so make sure to answer these questions while you read the lab PDF. The pGLO plasmid DNA and the original bacteria can be eliminated from providing the fluorescent source. Course Hero is not sponsored or endorsed by any college or university. Transformation Lab A Plasmid Discovery Labratory 6, AP Biology Abstract. The expansion puts pressure on the weakened cell wall, which then ruptures from the pressure. record your answers in your laboratory notebook. Cells that were not treated with the plasmid (LB/amp (-) pGLO and LB/amp/ara (-) pGLO plates) could not grow on ampicillin, whereas cells that were treated with the plasmid (LB/amp (+) pGLO and LB/amp/ara (+) pGLO plate) can grow on the LB/amp plate. If no bacterial colonies survive, then they were not ampicillin resistant (they were ampicillin sensitive). bacterial transformation lab answers quizlet pglo bacterial transformation lab answers virtual lab answer key Some results have been removed. Explain your answer. When they drop down to a lower energy state they emit a longer wavelength of visible fluorescent green light at 509 nm. An organism which reproduces quickly. What traits or characteristics should the organism have (or not have) to be sure it will not harm you or the environment? This transformation usually occurs within plasmids, which are small circular DNA molecules separate from its chromosome. 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