Introduction In this week’s laboratory period students had the opportunity to perform a common procedure preformed by many if not all microbiologists known as genetic transformation. Genetic transformation is the ability to move DNA into an organism and thereby altering its genotypic and genetic makeup (2). Genetic transformation has shown to have a wide variety of uses in many scientific studies. In agriculture‚ gene coding for traits such as frost‚ pest‚ or spoilage resistance have been genetically
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fischeri DNA and pGEM DNA was cut with restriction enzyme Sal I to prepare both for ligation. 3. Vibrio fischeri and pGEM DNA were ligated with T4 ligase. 4. E.coli DH5α competent cells were prepared for us so that transformation of the recombinant plasmids and the competent cells through heat shock transformation. 5. The transformation was then plated on LB/amp/X-gal plates. 6. The plates were screened for our target with ampicillin resistance‚ blue/white colony‚ then bioluminescence. 7. A bioluminescent
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plates were set up with agar in them for the bacteria to feed on and grow. Changes were then made to the bacteria. One plate was the control plate‚ having only the LB or agar for the bacteria and negative pGLO‚ which is the liquid not containing the plasmid. This is the plate that was compared with the three others in order to identify the changes. The second plate contained negative pGLO‚ LB‚ and ampicillin. This is to see whether or not the bacteria will become resistant to the ampicillin and grow
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see results fast‚ there is a commonly used non-virulent strain‚ and it is cheap to grow and does not require a lot of resources. A plasmid is what we are going to use to give the E. coli cells their extra genetic material. Plasmids are separate pieces of DNA that are not part of the circle of DNA that holds most of the genes in a genome (Brooker et al.‚ 2011). The plasmid‚ pGLO‚ is what we put inside the E. coli‚ it contains three genes: bla‚ araC‚ and GFP. The gene bla stands for β- lactamase‚ which
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Subcloning of fungal cDNA from pBK-CMW into a plasmid vector pUC19 using fungal gene CIH Introduction A plasmid is a circular‚ double stranded DNA molecule that replicates independently of the chromosome DNA within a cell.pUC19 is one of the most commonly plasmid cloning vector used due to its high copy replication number (approx. 100 copies per cell)‚ ampR (ampicillin resistance gene) andterminal fragment of β -galactosidase (lac Z). It is circular double stranded and it has 2686 base pair
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Malak Zomrawi 4/9/15 Bacterial Transformation I. Abstract In the lab‚ the purpose is to see if we could move genes using plasmid. As well as getting better understand of transformation methods using shock wave. To see the effects five trays are being used containing LB nutrient broth. The results showed that the LB‚ ampicillin‚ and arabinose with a positive pGLO had the most amount of growth compared to the other four trays. Although when there is arabinose there is no fluorescence‚ fluorescence
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reagent NB: • Using PLUS™ Reagent (Cat. No. 11514-015) enhances transfection performance in HUVEC cells. • The addition of antibiotics to media during transfection may result in cell death Transfection of HMECs Use this procedure to transfect plasmid DNA into HMECs cells in a 12-well format All amounts and volumes are given on a per well basis. 1. Cell density should be 50~80% confluent on the day of transfection (use the normal growth medium without antibiotics). 2. For each well of cells
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Report Backround: The plasmid pGLO contains an antibiotic-resistance gene‚ ampR‚ and the GFP gene is regulated by the control region of the ara operon. Ampicillin is an antibiotic that kills E. coli‚ so if E. coli‚ so if E. coli cells contain the ampicillin-resistance gene‚ the cells can survive exposure to ampicillin since the ampicillin-resistance gene encodes an enzyme that inactivates the antibiotic. Thus‚ transformed E. coli cells containing ampicillin-resistance plasmids can easily be selected
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The procedure was taken from “From Drosophila cDNA in E. coli plasmid to homologous human proteins” lab manual (4). - Colony Picking: Two E. coli colonies were grown on agar plates and treated with ampicillin. They contained the plasmid with genes for ampicillin resistance and Drosophila cDNA sequence. - Plasmid Isolation: We used the QuickLyse Miniprep Plasmid DNA purification systems to isolate the plasmid DNA. Indeed‚ the bacterial cells were removed from the liquid broth and were resuspended
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the vector or insert. a) A schematic of gene W is below. You want to clone all of gene W DNA into the p7012 vector. Give three different strategies that you could use to clone gene W into p7012‚ and obtain colonies that contain a recombinant plasmid. * Strategy 1 uses the restriction enzyme(s) ______Kpnl________ to cut the vector and restriction enzyme(s) ____Kpnl_________ to cut Gene W * Strategy 1 uses the restriction enzyme(s) _EcoRL and Sall_ to cut the vector and restriction
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