Arielle Fisher GFP Lab November 16‚ 2011 Figure 1 A. B. C. D. Figure 1. Confocal images at 400x magnification of HeLa cells to locate GFP activity. HeLa cells were (A) tagged with Green Fluorescent Protein (GFP) (B) labeled with GFP and 14.13 µl Dexamethasone (C) tagged with GFP fused to the glucocorticoid binding
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amount of protein‚ and percent yield and an increasing trend for specific activity and fold purification. These trends come about naturally when performing multiple purification steps. To determine the success of each purification step‚ the more important factors to look at are the total activity units‚ total protein amount‚ and percent yield. Looking at our purification table (table 7) our purifications were not very successful. While our total protein followed the normal decreasing trend‚ as
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To purify the protein in the cell lysate from lab 1 through nickel affinity chromatography. Protein purification should result in only one type of protein ideally‚ which is the protein of interest‚ wt-DHFR and mut-DHFR in this case. A pure protein allows for further analysis on the protein to be conducted‚ such as its concentration (Bradford assay)‚ its molecular weight‚ and its biological activity. 2. Overview of experiments Buffer Preparation Add the liquid sodium phosphate‚ solid sodium chloride
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The purpose of module E is to learn several DNA techniques in the lab including DNA purification with solubility and absorption‚ plasmid transfection of E.coli‚ colony screening by PCR and quantitative PCR. First part of the experiment E1 show the purification method of DNA through solubility. E. coli lysate mixed organic solvents to purify the DNA present in solution. First‚ the lysate was mixed with phenol/chloroform‚ then vortexed‚ and centrifuged. We extracted the aqueous layer and combined
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DNA can replicate inside the transformed E.coli DH5α cells‚ only successful transformed cells can produce the protein that is resistance to kanamycin‚ this allows for the selection of successful transformed cells. 2. Overview of experiments DNA purification Maintain the PCR volume of 50µL-100 µL. Pipet the elution buffer in the center of the PureLink® Spin Column (PSC) and perform an incubation. Add B2 to PCR reaction. Add this sample to the PSC. Centrifuge the PSC. Replace the PSC into the wash
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PURIFICATION OF LACTATE DEHYDROGENASE FROM CHICKEN MUSCLE TISSUE Abstract The enzyme lactate dehydrogenase (LDH) catalyzes the last step of anaerobic glycolysis that is important for the normal function of the body. Purification of LDH is essential to understand its structure and function. The purpose of this experiment was to extract and purify LDH enzyme from chicken muscle tissue using a variety of various. Analytical methods such as activity and protein assay were employed to determine
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Government-Furnished Property GFP What is Government-Furnished Property and Why do I care about it? Government-Furnished Property and You ▪ What is GFP? ▪ When do we provide GFP? ▪ Who is responsible for the GFP? ▪ The Government retains title to the GFP until it is properly disposed of as authorized by law and regulation. Government-Furnished Property Defined ▪ GFP is property in the possession of‚ or directly acquired by‚ the Government and subsequently furnished to the contractor for performance
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The Purity and Purification of Solids Recrystallization ABSTRACT In this experiment‚ impure adipic acid is purified though recrystallization. The impure adipic acid is dissolved in a solvent. After cooling it is ran through a vacuum filter to separate any moisture. The dry crystals are then weight for percent yield and the melting point range is taken to test the purity. There were two different trials ran in this experiment. The difference between the two was the addition of charcoal in
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What is water purification? Water purification generally means freeing water from any kind of impurity it contains‚ such as contaminants or micro organisms. Water purification is not a very one-sided process; the purification process contains many steps. The steps that need to be progressed depend on the kind of impurities that are found in the water. This can differ very much for different types of water. In which ways is polluted water treated? Settling Before the purification process begins some
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DNA extraction and purification is an essential tool in understanding the biological basis and significance of a eukaryotic cell. Its role is pivotal to numerous scientific applications ranging from basic science research to applied research. Hence‚ DNA is found in all living organisms. Fragaria x ananassa‚ also known as a strawberry has been widely used as one of the many biological models in studying DNA structures due to its accessibility. Strawberries are octoploid which contain large genomes
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