the cell competency‚ meaning that it will be able to take on additional DNA. This was done when the plasma was added. Materials: 1. 37 o C water bath 2. Ice 3. Sterile transfer pipette 4. Foam tube rack 5. Transformation solution (CaCl2) 6. pGLO plasmid 7. Sterile Inoculating loop 8. 2 - LB+amp plate 9. LB+amp+ara plate 10. LB plate 11. LB broth 12. E.coli starter plate culture Procedure: The test tubes were labael +pGLO and another –pGLO‚ and placed them on a test tube. Transfer 250uL of transformation
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incubation for at least 10 minutes. The cell competency preparation was carried out by the instructor in this experiment. Stage two was the uptake of DNA by competent cells. This stage was carried out by five groups‚ three performing plasmid pUC18 and two groups performing plasmid lux. Each group labeled two Eppendorf tubes‚ one was labeled
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DISCUSSION In this experiment‚ we will extract plasmid DNA that carrying pKan and pAmp from E. coli that have been cultured overnight in LB media containing antibiotic kanamycin and ampicillin respectively. So‚ the DNA that will be introduced later will make the bacteria resistance to antibiotic kanamycin or ampicillin. A plasmid is a small‚ circular‚ double stranded DNA molecules and cloning vector that are widely used for recombinant DNA technology. It can be physically separated from chromosomal
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Material and methods Hosts ‚ plasmids and chemicals E. coli TOP10 strain was used for cloning and proliferation. E.coli BL21 DE3 was used as expression host cell. The pET 28a(+) plasmid was employed for gene expression. All chemicals were obtained from Merck Company (Germany). Codon optimization and gene synthesis Sequence encoding V-domain was obtained from Swiss-port‚ Uniprot KB and National Center for Biotechnology Information (NCBI) databases. 6x His-tag sequence was placed at N-terminal
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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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Title: E. Coli Transformation with a Plasmid DNA Containing the GFP Gene Introduction: Bacterial transformation is the process of bacteria taking in and expressing exogenous DNA. This has led to many other discoveries. In order for bacterial transformation to occur the bacteria must be in a certain physical state to be able to take in DNA. This is called competency and it allows the cell membrane to be permeable so DNA can pass through. Currently researchers are studying the transformation
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buffer (pH 6‚0) overnight at room temperature‚ frozen in liquid nitrogen and stored at -80 ˚C after 0.22 μm pore membrane filtration. Evaluation of DNA-protein interaction by gel retardation assay Protein-DNA complexes were formed using 1 µg of plasmid pTriEx-1.1 Hygro at different pDNA: protein molar ratios (1:100‚ 1:200‚ 1:500‚ 1:1000‚ 1:2000‚ 1:8000) during 10 minutes. The increasing molar ratios were assessed by gel retardation assay on a 0.8% agarose gel and visualized by ethidium bromide staining
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BIO 219 Group 1 Section 66 October 19‚ 2012 The extraction of purified DNA from A. fischeri by restriction digestion using Sal I enzyme and pGEM for shotgun cloning Introduction: The ultimate goal of this experiment is to isolate the lux operon‚ a targeted piece of DNA that causes bioluminescence‚ from Aliivibrio fischeri and insert it into the DNA of Escherichia coli in order to make it glow. A. fischeri is a gram-negative bacteria which participates in a symbiotic relationship with
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1 THE LURIA-‐DELBRÜCK EXPERIMENT The Luria-Delbrück Experiment Henry Paige University College Freiburg 2 THE LURIA-‐DELBRÜCK EXPERIMENT Abstract This paper discusses Luria and Delbrück’s 1943 Fluctuation Test. Historical background is first addressed to contextualize the experiment and define the theories‚ ideas‚ and discussions relevant at the time of the experiment. A brief summary of biologists’ knowledge and attitudes towards bacteria is explained through contemporary quotes. The two main
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In the experiment involving varying pH levels‚ E. faecalis‚ S. epidermidis‚ L. lactis‚ and L. casei were subjected to different pH levels and then were allowed to incubate in order to determine the minimum‚ maximum‚ and optimum pH levels for growth of specific bacterial species. It was found that lower pH levels between 2 and 4 inhibited or promoted little to no growth for E. faecalis‚ S. epidermidis‚ and L. casei. It is evident that these bacteria were resistant to alkaline environments‚ however
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