EXPERIMENT NO. 15 PROTEIN CHARACTERIZATION BY ELECTROPHORESIS Abstract The molecular weights of protein extracts were assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Two sets of four protein samples‚ standard bovine serum albumin (BSA)‚ invertase‚ egg albumin‚ and casein‚ were prepared; one set containing β-mercaptoethanol (BME) while the other did not. These were then analyzed through SDS-PAGE with 12.5% resolving gel‚ prepared using 2 M Tris-HCl at pH 8
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their absorbance readings will determine if they are haemoglobin or cytochrome c. Background research: A technique widely used by scientists for over 30 years gel filtration chromatography purifies biological samples according to their molecular size and weight. The procedure involves a column of beads‚ containing within various pores which allows the heaviest molecule to penetrate through the column and eluting first‚ and then decreasing in size and weight in order. Globular proteins have
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AGAROSE GEL ELECTROPHORESIS Gel electrophoresis is a widely used technique for the analysis of nucleic acids and proteins. Most every molecular biology research laboratory routinely uses agarose gel electrophoresis for the preparation and analysis of DNA. We will be using agarose gel electrophoresis to determine the presence and size of PCR products. Background: Electrophoresis is a method of separating substances based on the rate of movement while under the influence of an electric field
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the purification and size determination of various proteins and DNA fragments. In order to do this‚ a polyacrylamide gel will be prepared and placed in a buffer-containing gel electrophoresis apparatus. Next‚ an aliquot of acid phosphatase and a molecular weight marker (Composed of Phosphorylase B‚ bovine serum albumin‚ ovalbumin‚ and carbonic anhydrase) will be placed into separate wells within the gel‚ and the apparatus will be connected to a constant voltage source (175 V) for an allotted period
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Lesley F. Biology 902 November 7‚ 2013 18.1 (1-3) 1a. What are the two goals of systematics? The goal of systematics is to organize living things into groups that have biological meaning. (Taxa or Taxon) 1b. Why do common names of organisms – like daisy or mountain lion – often cause problems for scientists? The common names of organism often cause problems for scientist because common names can change meaning among languages and from place to place. 1c. The scientific
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IPTG induction‚ salting out‚ thermostable polymerase Background: Taq polymerase is a thermostable enzyme essential in the PCR reaction that is isolated from recombinant E. coli. Results: Taq polymerase was isolated‚ the sample was not pure Taq‚ molecular weight found to be 114kDa. Conclusion: The methods isolated taq but an improvement of cation exchange and increase heat denaturation should be added. Significance: An improved method of Taq isolation offers a valuable resource for PCR. SUMMARY The
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Page 1 Analysis of transcriptional regulation of the arabinose operon promoter using expression from a green fluorescent protein encoding reporter gene + + KIRA FERNANDEZ ‚ CLAIRE MARKEY * ‚ JESSE PIERATTI + Department of Biological Sciences ‚ and Department of Nutrition and Dietetics*‚ Messiah College‚ Mechanicsburg‚ PA 17055 Page 2 ABSTRACT This study investigates gene regulation and how environmental arabinose and/or glucose can interact with geno
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Introduction: Escherichia coli (E.coli) is a commensal-pathogenic organism which include a wide range of strains. There are several advanced molecular-genomic technologies that are capable for detecting and identifying different strains of E.coli. But‚ ERIC-PCR technique is a quick and cost effective method for determining individual strains via demonstration of strain specific fingerprint bands. Therefore‚ the ERIC-PCR technique was used to determine the isolated strains of E.coli from different
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Gel Filtration Gel filtration is a non-adsorptive chromatography technique that separates molecules on the basis of molecular size. Desalting and buffer exchange are two special examples of gel filtration that are widely used in many downstream bioprocesses. Desalting is used to completely remove or lower the concentration of salt or other low molecular weight components in the sample while buffer exchange replaces the sample buffer with a new buffer. Gel filtration is one of the easiest chromatography
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WESTERN BLOTTING (Written Report) I. PRINCIPLE Western blotting is a method for identifying a specific protein in a complex mixture and simultaneously determining its molecular weight. In Western blotting‚ proteins are electrophoresed into a gel‚ as the proteins migrate through the gel they are separated based upon size and charge. Characteristically‚ smaller proteins migrate through the gel faster than larger proteins. Sufficiently separated proteins in an SDS-PAGE can be transferred to a
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