E. Coli Transformation with Plasmid (pGal)‚ pGal Isolation‚ and Analysis of Plasmid DNA Felicia Osadi Bio 22 April 20‚ 2012 Transformation = group 10 Plasmid = group 7 RFLP = group 1 RESULTS Table I. Plasmid Transformation of E. Coli. Plate # | Agar plate | Type | Result | 1 | X-gal | Control | Extensive lawn growth | 2 | Ampr / X-gal | Control | Clear no bacterial growth | 3 | Ampr / X-gal | Transformation | 1 blue colony | Transformation efficiency = 1 transformants
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Wintersemester / page 1 Experiment 22 Isolation of plasmid-DNA from bacteria and PCR Advisor Konrad Egli: kegli@botinst.unizh.ch Reading Chapters in BBOM 10th: 10.8 BBOM : Madigan M.T.‚ J.M. Martinko and J. Parker: "Brock - Biology of Microorganisms"‚ 10th Edition (2003)‚ Prentice Hall. Objectives Background • Isolation of plasmid-DNA from different bacteria clones • Handling of bacteria clones • PCR-experiment The typical plasmid is a circular double-standed DNA molecule less
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Bacterial Transformation Lab Introduction: In this experiment we transformed a strain of E. Coli bacteria without antibiotic resistance with plasmid DNA. This plasmid produces a fluorescent green glow under black light due to the gfp(green fluorescent protein) as well as antibiotic resistance. E. Coli cells will be plated on an agar medium‚ some with and some without the antibiotic ampicillin. Only bacterial cells that contain the plasmid will survive the ampicillin and produce the green glow
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RNA translates it into protein and expressing it as a trait. Recombinant plasmids are when DNA fragments are inserted into a plasmid vector. The recognition site is where the plasmid gets cut by the restriction enzyme which is an enzyme that cuts a DNA molecule. Recombinant DNA is the DNA being inserted into the E.Coli. The purpose of the lab is to see if we can make the E.Coli glow and resistant to ampicillin. In the lab we were transforming a north american jellyfish by the name Aequorea victoria
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genetic transformation in bacteria (E. Coli). More specifically‚ a previously prepared pGLO plasmid--which consisted of the gene to be cloned--was used to transform non-pathogenic bacteria. The pGLO plasmid contained a gene for the Green Fluorescent Protein (GFP) from a bioluminescent jellyfish and a gene for resistance to ampicillin‚ an antibiotic. Essentially‚ we wanted to determine the conditions of the bacteria that would glow. Our hypothesis was that the transformed solution with no plasmid DNA
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November 25‚ 2012 The Effect of the pGLO Plasmid on Genetic Transformation of E.coli by Heat Shock Introduction Genetic transformation is the genetic alteration of a cell resulting from the direct uptake‚ incorporation and expression of exogenous genetic material l(exogenous DNA) from its surroundings and taken up through the cell membranes. This was first demonstrated in 1928 by Bacteriologist Frederick Griffith (Lederberg 2000).A plasmid is a small circular piece of DNA that contains important
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uses of plasmids in G.M. experimentation. Plasmids are extrachromosomal genetic elements found in a variety of bacterial species. They are double stranded; autonomously replicating‚ supercoiled‚ covalently closed circular (ccc) DNA molecules that range in size from 1 kb to greater than 200 kb. Often‚ plasmids contain genes coding for enzymes that‚ under certain circumstances‚ are advantageous to the bacterial host (Table 1). Table 1. Some of the phenotypes conferred by different plasmids that
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LSM1102 Lab Report Introduction Transformation is a process which involves plasmid DNA being bound to the cell surface and the subsequent uptake of DNA by the cell (Panja et al.‚ 2008). For artificial transformation of E. coli cells with plasmids‚ plasmid DNA has to be extracted from bacterial cells using the High-Speed Plasmid Mini Kit‚ which is then mixed with competent E. coli cells followed by heat shock and the streaking of transformed cells on two different types of agar plate (LB and LB+ampicillin)
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I. Title Identification of an Unknown Plasmid In this experiment‚ we determined the phenotypic capability of an unknown plasmid along with its size. With the use of gel electrophoresis‚ we analyzed the gel photograph by using a standard DNA marker‚ Lambda HindIII‚ and came to a conclusion based on our results. II. Abstract Two experiments were done to identify an unknown plasmid. The success of these experiments came from the use of modern day technology involving gel electrophoresis
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Glowing Transformations Abstract In this experiment‚ the idea is to become familiar with the transformation of cells. A well thought out procedure‚ involving a heat shock procedure‚ a good antibiotic‚ an inducer known as arabinose to show the newly expressed DNA by a visible fluorescent glow‚ and a stable control group is what contributes to this experiments thoroughness. It is predicted that the four agar plates will all yield different forms of growth‚ with different coloration and colony
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