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‚ with elution 3 containing
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CHEM 2204 Chromatography Lab by wyk.wong » Fri Jul 11‚ 2014 10:25 am Results and Calculations Rf values Rf=(Distance moved by the spot (cm))/(Distance moved by the solvent front (cm)) Toluene: Rf=2 cm/3.8 cm=0.53 (Fluorenone) Rf=1.1 cm/3.8 cm=0.29 (Fluorene) Hexane: Rf=1.8 cm/2.2 cm=0.82 (Fluorene) Rf=0 cm/2.2 cm=0 (Fluorene Table 1: Experimental IR peaks compared to literature IR peaks for fluorenone Functional group Experimental peak (cm-1) Literature peak (cm-1) C-H 3010.5 3013
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When the actual FD&C dyes in the green Kool-Aid were revealed‚ it was discovered that the initial assumption that Yellow 5 and Blue 1 are in the Kool-Aid was correct. However‚ solely based upon the Rf values‚ the dyes in the green Kool-Aid are Red 40 and Yellow 6 as those are closet Rf value to the numeric data collected and calculated from the Kool-Aid chromatogram. However‚ the chromatography paper in both trials display that the dyes in Kool-Aid are a form of yellow and a form of blue because
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Liquid Chromatography Purpose: The purpose of this lab was to separate substances based on their polarity by using liquid chromatography. Data Table: Red Dye Blue Dye Run#1 Run#2 Run#3 Run#1 Run#2 Run#3 Start of Band(mL) 1.50 2.20 1.00 2.70 3.00 2.00 End of Band(mL) 2.70 3.00 2.00 6.40 5.50 6.00 Beaker Eluant Observations 1 H2O White powder 2 5%isopropyl Red powder 3 28%isopropyl Blue powder 4 70%isopropyl Oily residual Calculations: W = Vend – Vstart
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Post Lab #4- Column Chromatography Organic Chem 3418-2 March 3‚ 2011 Theoretical Background- The fluorene and fluorenone mixture was separated by first dissolving the mixture in heptane. Since “like dissolves like”‚ fluorene dissolves with the non-polar heptane and the polar fluorenone dissolves in the polar ethyl acetate solvent. This phenomenon was illustrated in class before the experiment‚ when it was pointed out why water will not dissolve fluorene‚ fluorenone‚ or transstilbene
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Chromatography – Analyzing Analgesics by TLC and Isolation of β-Carotene by Column Chromatography Introduction/Background: Flavonoids are an important group of additives that can be defined as pure substances either natural‚ extracted from raw materials or synthetic. Chromatography is the separation of two or more compounds or ions caused by their molecular interactions with two phases – one moving and one stationary (Weldegirma 2012). Three types of chromatography are used
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Encyclopædia Britannica Paper chromatography is used to separate "dissolved chemical substances". An example of dissolved chemical substances could be the pigments that make up a specific color‚ for example in a marker. When a chromatography paper experiment is done on a black marker‚ the experiment tends to separate all the different pigments that make up the color. In this experiment‚ the substance investigated was a black water soluble Giotto marker pen. Paper chromatography has two phases‚ the
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Gas Chromatography Analysis of Product Mixtures Gas Chromatography Guidelines‚ Handout. Introduction Gas chromatography is a technique used to analyze chemical compounds that can be vaporized and separated in a gas phase column. Once separated‚ the analyzed substance is passed through a detector and data is obtained. The samples that we are going to analyze are: the EtOAc from Simple distillation‚ the Fraction 1‚ Fraction 2‚ and Fraction 3 from the Fractional Distillation. Experimental Procedure
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org/wiki/Chromatography http://ull.chemistry.uakron.edu/analytical/Chromatography/ http://orgchem.colorado.edu/hndbksupport/TLC/TLC.html this is for TLC – similar to paper http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/C/Chromatography_paper.html http://jchemed.chem.wisc.edu/JCESoft/Programs/CPL/Sample/modules/paprchrom/paprchromdesc.htm http://jchemed.chem.wisc.edu/JCESoft/Programs/CPL/Sample/modules/paprchrom/paprchromdesc.htm This site shows the colors of many of the food colorings
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The purpose of column chromatography is to separate one compound from another by the differing polarities in the compound. This technique is important in organic chemistry because certain experiments may require the separation of compounds in solution to be used for chemical synthesis and/or analysis of a particular desired product by isolating it. Fluorene is to be separated from 9-fluorenone using the technique of column chromatography. The success of separation and purity is determined with analyzation
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