1. Plasmids Plasmids are closed circular‚ double-stranded‚ extrachromosomal DNA molecules which occur naturally in bacteria‚ yeast‚ and some higher eukaryotic cells‚ and exist in a parasitic or symbiotic relationship with their host cell (Lodish et al.‚ 2000) The main application of plasmids is as cloning vectors in gene cloning. In gene cloning‚ a fragment of DNA‚ containing the gene to be cloned is inserted into a circular molecule called the “vector” to produce recombinant DNA molecule. Plasmids
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to be present using a restriction enzyme digest; after which the cloned genes were inserted into the bacteria E. coli through the transformation process consisting of a restriction enzyme digest and ligation. Colonies of the bacteria were grown; plasmids from each colony were purified‚ and then
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to successfully infiltrate E. coli bacterial cells with a pARA-R plasmid that is antibiotic resistant and has the rfp gene‚ or red fluorescent protein. This can be verified if the E. coli obtains the characteristics of the plasmid when it enters. To start‚ three Petri plates containing agar are needed. On each plate there is a control group and a treatment group; the treatment group being the one with the plasmid. Before the plasmid is put with the E. coli‚ first the bacteria are “stressed out” by
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electrophoresis. Lane 7: 10 µL of ladder. We began our goal of combining DNA by obtaining the desired samples of plasmid DNA from a liquid bacteria culture. Plasmids are circular pieces of DNA‚ which can be found in bacteria. Plasmids are often used in the manipulation of genes. After carefully following a mini-prep procedure‚ we were left with samples of pAMP and pKAN plasmids. pAMP is a bacterial plasmid‚ which contains the gene ampR. The ampR gene codes for a protein that breaks down the
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histidine-tagged pbluescript in a forward orientation potentially allows isolation of protein via Affinity Chromatography or Chromatin Immunoprecipitation therefore its role‚ effects and targets in the genome can be established. Resultant Recombinant plasmids in this experiment had multiple inserts leading to inconclusive orientation of the inserts; however this can be tackled by Sanger or Maxam/Gilberts sequencing. Introduction The capacity to segregate and amplify individual genes from an intricate
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PLASMID DNA ISOLATION‚ RESTRICTION ENZUME DISGESTION AND AGAROSE GEL ELECTRIPHORESIS Abstract: The gel is covered with an ion- containing buffer‚ such as (TAE)‚ that controls the pH of the system and conducts electricity. overall DNA concentration was lower than expected. Using agarose gel electrophoresis is to separate and visualize the DNA fragment‚ which is produced by restriction enzymes . Introduction: The purpose of this experiment is to measure the size of the fragments of DNA and separate
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toward Ampicillin due to Plasmid and DNA Consumption U34 Abstract During the ampicillin experiment the ability to transform cells to make them adaptable to their environment was studied. The E.coli bacterial cell was used in order to observe how its DNA was able to change and develop immunity towards ampicillin. In order for this change to occur the use of several plasmids was needed. The plasmids used in this experiment were pUC18 and the lux plasmid. The E.coli cells that
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Chapter 9 DNA-Based Information Technologies Multiple Choice Questions 1. DNA cloning: the basics Page: 307 Difficulty: 1 Ans: C Restriction enzymes: A) act at the membrane to restrict the passage of certain molecules into the cell. B) are highly specialized ribonucleases that degrade mRNA soon after its synthesis. C) are sequence-specific DNA endonucleases. D) are very specific proteases that cleave peptides at only certain sequences. E) catalyze the
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purpose of this experiment is to determine the effects of the addition of a plasmid to a bacterial cell. The bacteria E. Coli was separated into two groups: one where the pGLO plasmid was added to the bacteria‚ which contains the genes of fluorescence and resistance to antibiotics‚ and the other lacking the plasmid. The two groups then placed in agar plates simulating different environments: the bacteria lacking the pGLO plasmid was subjected to an environment solely contaiing nutrients and another containing
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has increasingly come to the forefront is DNA vaccination. DNA vaccines are bacterial plasmids that encode the polypeptide sequence
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