P lab 10 bacterial transformationpglo system questions. This shows that protein is present in the plate and it fluorescing due to it. When lab is complete, collect all petri dishes, open, and immerse in a 10% bleach. I can successfully transform the bacteria and investigate the heat factor in bacterial transformation. Coli with pglo plasmids, a lab day one transformation background. In bacteria, a small circular piece of dna known as a plasmid table 1, transfers genetic information between. The detailed lab protocol can be found in the student manual. However, with this specific lab especially i found the writing of the lab report was just as educational if not more than the actual lab. See instructions in teachers manual for proper disposal. Tn5 transposon tools technical manual epicentre biotechnologies.
Pglo transformation lab ap lab 7 capital high school. Introduction we did the experiment to learn about the transformation of the gfpgene. Transformation is a process of transferring genetic information from one organism to another. Some terms i dont understand and how they correlate to each other.
Duplication of any part of this document is permitted for classroom use only. As we learned at the beginning of unit 3, dna is found in the form of a double helix, meaning that it has two strands, and is composed of nucleotides, with each nucleotide itself being composed of a deoxyribose sugar that is bonded by a. The purpose of this lab was to genetically modify e. However, with this specific lab especially i found the writing of the lab report was just as educational if not more than the actual. Pdf bacterial transformation and the origins of epidemics in the. The active chromophore of gfp is comprised of three adjacent amino acids in the primary amino acid chain. When transformation is complete, and we insert the plasmid into a bacteria cell, the cell will express the jellyfish gene. The hypothesis was supported by this experiment because in the presence of the gfp gene the bacteria grew and glowed. When transformation is complete, and we insert the plasmid into a. These plates contain the bacterial nutrient medium. I can successfully transform the bacteria and investigate the. What i learned from this labimpressions of the pglo lab.
Effect of ampicillin on bacterial growth lab open inquiry. Student manual pglo transformation lesson 1 introduction to transformation in this lab you will perform a procedure known as genetic transformation. Bacterial transformation with green fluorescent protein pglo genetic engineering is an umbrella term that encompasses many different techniques for moving dna between different organisms. View notes pglo lab from biol 6b at deanza college. Connor lauffenburger 317 pglo transformation lab report i introduction the purpose of this experiment was to show the genetic transformation of e. The video includes a walk through of the protocol as well as background information on plasmids, operons and. For technical service call your local biorad office or in the u. Jan 31, 20 as you can see, we were testing with several different variables, including pglo presence, ampicillin presence, and arabinose presence. Essay on pglo transformation lab report 1486 words. This new genetic information often provides the organism with a new trait. Add nothing, 1 microliter of pglo dna, and 1 microliter of tn5 mutagenesis pglo reaction to each tube respectively.
Read this lab and be ready to get down the exercisings. Ultimately what we learned from this lab was the workings of genetic transformation. In lab groups, we genetically engineered the bacteria with pglo plasmids containing specific codes for gfp green fluorescent protein and ampicillin, an antibiotic, resistance. In this lab you will perform a procedure known as genetic transformation. Introduction to explain the pglo transformation lab, we must first go all the way back to dna structure. To develop newly acquired skills in the laboratory. We just completed the pglo laboratory and i have no clue of what we are doing, and how the lab exactly works the technicalities of it. When we shined the uv light onto the negative pglo plasmid dna we did not see anything significant happen and the colonies were unaffected. Learn vocabulary, terms, and more with flashcards, games, and other study tools. When we shined the uv light on the positive pglo plasmid dna we saw the colonies in the plates fluoresce a green colour. Could someone be so kind enough as to explain it to me please.
All books are in clear copy here, and all files are secure so dont worry about it. Specify the undermentioned footings but do non manus in. The plasmid contains several reporter genes, most notably the green fluorescent protein gfp and the ampicillin resistance gene. In this laboratory investigation, plasmids containing fragments of foreign dna will be used to transform escherichia coli cells, conferring antibiotic ampicillin resistance, as. Bacterial transformation lab by lindsay young on prezi. Bacterial transformation the purpose of this lab is to demonstrate visible changes in e. The recombinant pglo plasmid contains the gene for green fluorescent protein gfp and a. There is another important gene on this pglo plasmidan antibiotic resistant gene that. Mar 02, 2018 here i discuss a popular bacterial transformation lab using the pglo plasmid. Download pglo transformation lab answer key pdf book pdf free download link or read online here in pdf. The progression from dna rna protein trait is shown with the bacteria e.
This site is like a library, you could find million book here by using search box in the header. Read about the control of cistron look on pages 353356 and about transmutation on page 348 of the text edition. The source of this gene is the bioluminescent jelly fish aequorea victoria, and the gfp causes the. This plasmid contains the original dna as well as a gene from a jellyfish. Call 18004biorad 18004246723 pglo arac gfp bla ori. Download the complete instruction manual for the bacterial transformation. You will be using a plasmid called pglo p plasmid because the gene being carried on is the code for the fluorescent protein. The source of this gene is the bioluminescent jelly fish aequorea victoria, and the. To get acquainted with bacterial transformation applications in society. Biotechnology explorer bacterial transformation the pglo system catalog number 1660003edu.
The lbamp pglo plate will have no growth, the lb pglo plate will have a lawn of growth meaning colonies covering. Transformation is the process by which an organism acquires and expresses a whole new gene. Student manual pglo transformation lesson 1 introduction to. This comparison shows that genetic transformation produces bacterial colonies that can grow on ampicillin due to the uptake of the pglo plasmid and the expression of the ampicillin resistance gene. Introduction abstract our alternate hypothesis for this experiment will be that the pglo dna will incorporate in the e. Transformation of cells is a widely used and versatile tool in genetic.
Bacterial transformation with pglo overview transformation modification of a bacterium by the uptake and incorporation of exogenous dna determine the transformation efficiency of the competent cells. The gene for gfp can be switched on in transformed cells simply by. Bacterial transformation the pglo system catalog number 1660003edu. Investigation 8 bacterial plasmidbased genetic transformation, enables students to. Coli with pglo plasmids, a lab book pdf free download link book now. Heat shock the tubes at 42 degrees for exactly 90 seconds and return to ice immediately for 2 minutes. Our lab is performing transformation we will take a plasmid that has be recombined into a piece of recombinant dna. Genetic engineering is the directed transfer of a gene, or piece of dna, into a cell typically a 6. Pdf frederick griffith 18791941 was an english bacteriologist at the. Bacterial transformation serves as an essential basis to a number of dna technologies. Its transformed with the help of pgloplasmid into e. This site is like a library, you could find million book here by using. Remember that a gene is a piece of dna which provides the instructions for making codes for a protein.
Therefore, the control plate was the one without all of these i. Gently mix tubes by tapping them and place on ice for 15 minutes. This site is like a library, you could find million book here by. The general process by which foreign dna is introduced into a cell is called transformation. The manual includes the teachers guide, student manual, and graphic quick guide. The pglo system also allows an additional experiment involving purification of. Read online pglo transformation lab book pdf free download link book now. Competent cells have the ability to take up dna molecules from the environment. Remember that a gene is a piece of dna that provides the instructions for making coding for a protein that gives an organism a particular trait. C hot water, incubated and then cooled down on ice. Read online pglo transformation lab answer key pdf book pdf free download link book now.
Analysis of results the goal of data analysis for this investigation is to determine if genetic. Bacterial transformation with pglo essay bacterial transformation with pglo essay bacterial transformation with pglo essay. Genetic transformation using bacteria and the pglo plasmid biology libretexts. Feb 23, 2016 completing the pglo experiment was really useful for learning about the processes of bacterial transformation and how to properly follow the lab procedure. Pglo transformation lab answer key pdf pdf book manual. Introduction to transformation in this lab you will perform a procedure known as a genetic transformation. This series of videos illustrates the steps required to pour agar plates and streak them with e. The pglo plasmid is an engineered plasmid used in biotechnology as a vector for creating genetically modified organisms. Presented in this way, griffiths experiment on bacterial transformation also ceases to appear. Transform bacteria using the pglo lab procedures big ideas. Essay on pglo transformation lab report 1486 words bartleby. The pglo system the recombinant pglo plasmid contains the gene for green fluorescent protein gfp and a gene for resistance to the antibiotic ampicillin.
Abstract plasmids are pieces of circular dna that are in bacteria which can code for genes that can cause the bacteria to have extra features. With the pglo plasmid this extra feature causes bacteria to glow from the green fluorescent protein that was inserted and also has resistance to ampicillin from the betalactamase protein. Completing the pglo experiment was really useful for learning about the processes of bacterial transformation and how to properly follow the lab procedure. Transformation of pglo into escherichia coli author. Gfp was isolated from the jelly fish aequorea victoria. Genetic transformation is the process by which an organism acquires and expresses a new gene. The lab protocol can be conceptualized as four major steps. The pglo plasmid showing positions of genes of interest. The bacteria will then exhibit a glow in the dark property because it incorporates this. It occurs when a cell takes up takes inside and expresses a new piece of genetic materialdna. This instruction manual, and the experiments outlined within it, are designed. The pglo bacterial transformation lab is based off the molecular biology ap biology lab involving transformation. Bacterial transformation is a really easy way to transform due to the fact that it is single cell.
Bing ap biology 6 february 20 pglo transformation lab introduction. Download pglo transformation lab book pdf free download link or read online here in pdf. Here i discuss a popular bacterial transformation lab using the pglo plasmid. As you can see, we were testing with several different variables, including pglo presence, ampicillin presence, and arabinose presence. The arabinose is a promoter aided with the glow of the pglo gene.
This shock is important for the success of the transformation. The purpose of the pglo lab is to insert a plamid into the dna of a bacteria such as li. Genetic transformation of the green florescent protein gfp gene into the bacterial cells of escherichia coli by use of the heat shock method of transformation. Mar 27, 2014 label the three tubes, no dna, pglo wt, and pglo mutant. Genetic transformation using bacteria and the pglo.
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