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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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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This experiment tested the growth of E.coli with inserted plasmid on an agar plate with Ampicillin. One colony of E.coli resistant to Ampicillin was grown during this experiment. The overall goal of the experiment was to successfully grow E.coli on the agar plate‚ which would show that the plasmid had been effectively inserted into the bacteria’s genes. This experiment helped students understand how plasmids were inserted into bacteria and used in real life situations. It also showed how the bacteria’s
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The pGLO plasmid‚ which has the bla and GFP gene‚ was developed so it can operate like the arabinose operon and be able to transcribe genes in the presence of arabinose sugar. In this plasmid‚ the cluster of genes is regulated by the spontaneous on/off element by a single promoter‚ which is dependent on the DNA binding protein araC. This protein is at the binding site for RNA polymerase at the beginning of the operon‚ so when arabinose is present (like in one of the experimental plates)‚ it is taken
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Molecular Biology Lab Report Payton Jackson Introduction In this lab‚ I am going to use antibiotic-resistance plasmids to transform Escherichia coli. Materials For this lab you will need the following: LB Agar Petri dishes Beakers Test tubes CaCl2 solution Sensitive E. coli (-ampR) amp plasmids ampicillin -amp cells Water bath to heat shock cells A freezer to incubate cells Process Step 1: Wash hands and sanitize lab setting. This will prevent anything reacting with a
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using calcium chloride/heat shock. This is done by incorporating the plasmids into chemically competent cells that were made permeable by the calcium chloride solution and heat shock. In 1928‚ Frederick Griffith‚ a physician from London‚ was he first person to experiment with bacterial transformation. He permanently transformed a safe‚ nonpathogenic bacterial strain of pneumococcus into a deadly pathogenic strain. [1] Plasmids are small circular autonomously replicating pieces of DNA that can be
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