The Expression and Purification of Recombinant Green Fluorescent Protein (rGFP) From E. coli strain‚ BL21(DE3)‚ Using Ni2+-Agarose Affinity Chromatography Abstract: The purpose of these series of experiments was to express and purify recombinant Green Fluorescent Protein (rGFP) from the E. coli strain‚ BL21(DE3) by beginning with its purification via a Ni2+-agarose affinity chromatography column. The His6 tag of the rGFP bound to the Ni2+-agarose column and washes and elutions were obtained
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unknown compound. In this experiment‚ chromatography and titration were performed to identify the unknown amino acid. Within experimental error‚ the results were consistent with the reference literature cited in this report. Experimental Thin Layer Chromatography The amino acid standards used in this experiment were Alanine‚ Glycine‚ Serine‚ and Histidine. These standards and the unknown were separated by performing a method of chromatography. Thin layer chromatography (TLC) was performed by using a mobile
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Thin Layer Chromatography/ Paper Chromatography Lab # 10 11/26 Sample # 32 for TLC Sample # 1 for Paper Introduction: Chromatography is one of the most important separation techniques used in all fields of chemistry ranging from analytical chemists to pharmacists. The understanding of how chromatography works and how to operate instruments used to carry out the procedures is an important lab technique to learn. Experiment: Thin Layer Chromatography. Lab #10-1 Paper Chromatography
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favorite candies? Chromatography can solve that for you. Chromatography is a separation technique used by scientists for separating both organic and inorganic compounds. There are four different types of chromatography: thin layer‚ liquid‚ gas‚ and paper‚ but for this lab paper chromatography will be used. Who invented chromatography? A Russian botanist named Mikhail Semyonovich Tsvet invented chromatography in 1901 while doing research on plant pigments. Why is chromatography so important? This
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Agarose gel electrophoresis is a technique used in the laboratory to separate macromolecules such as nucleic acids and proteins. Electrophoresis can take a mixture of macromolecules of different molecular weights‚ shapes‚ and various electrical charges to determine all the various compounds in the mixture and allowing for further purification that can aid in details of individual elements of the mixture being studied. Agarose gel electrophoresis is a very important technique used in the field of
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It was concluded that the product created was paracetamol when a thin layer chromatography test was performed and observed under ultraviolet light (which is discussed further down the page). An unknown substance was treated with acetic anhydrate and resulted with paracetamol. A compound that behaves in this matter is 4-aminophenol and is widespread in the industrial production of this drug. Upon these observations was the build up of “Chemical A” decided. When 4-aminophenol is t treated with
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other materials may be stored at room temperature (approximately 25°C). Use and Lab Safety: The materials supplied are for use with the method described in this kit only. Use of this kit presumes and requires prior knowledge of basic methods of gel electrophoresis and staining of DNA. Individuals should use this kit only in accordance with prudent laboratory safety precautions and under the supervision of a person familiar with such precautions. Use of this kit by unsupervised or improperly
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Chromatography • • • Separation based on polarity of compounds Two potential phases for a compound to exist in: mobile and stationary Partitioning of compounds between mobile phase and stationary phase occurs: o Compounds that are less polar move more in the mobile phase‚ those that are more polar “stick” more on the stationary phase o These polarity differences cause compounds move at different rates and therefore can be separated 1. Mobile Phase: the phase the moves; can be gas or
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The Isolation‚ purification and identification of Proteins assayed From Bovine Liver Using SDS Gel Electrophoresis‚ Mass Spectroscopy and Western Blotting Abstract The purpose of the experiment was to isolate and recognize varying protein solubilization and assaying methods by using bovine liver protein. The experiment implicated the impact of different types of solvents like ethanol‚ water‚ PBS‚ PBS+1% Triton x-100‚ and PBS+2% SDS on protein solubilization. Bradford and Ghosh/Dumbroff methods
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Discussion Using the Thin Layer Chromatography experiment‚ caffeine was found to be the most polar due to the fact that it stayed closer to the stationary phase. Caffeine contains four amine groups that are extremely polar as a result of the hydrogen bond and amide functional group. Acetaminophen was found to be the second most polar analgesic drug tested. Acetaminophen contains a polar alcohol group on one side and amide group on the other but also includes non-polar functional groups that consisted
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