title: Applied spectrophotometry: analysis of a biochemical mixture.
abstract: Spectrophotometric analysis is essential for determining biomolecule concentration of a solution and is employed ubiquitously in biochemistry and molecular biology. The application of the Beer-Lambert-Bouguer Lawis routinely used to determine the concentration of DNA, RNA or protein. There is however a significant difference in determining the concentration of a given species (RNA, DNA, protein) in isolation (a contrived circumstance) as opposed to determining that concentration in the presence of other species (a more realistic situation). To present the student with a more realistic laboratory experience and also to fill a hole that we believe exists in student experience prior to reaching a biochemistry course, we have devised a three week laboratory experience designed so that students learn to: connect laboratory practice with theory, apply the Beer-Lambert-Bougert Law to biochemical analyses, demonstrate the utility and limitations of example quantitative colorimetric assays, demonstrate the utility and limitations of UV analyses for biomolecules, develop strategies for analysis of a solution of unknown biomolecular composition, use digital micropipettors to make accurate and precise measurements, and apply graphing software.
authors.affiliation: Department of Chemistry and Biochemistry, Bloomsburg University of Pennsylvania, Bloomsburg, PA 17815, USA. mpugh@bloomu.edu
authors.forename: Toni A
authors.initials: TA
authors.lastname: Trumbo
authors.forename: Emeric
authors.initials: E
authors.lastname: Schultz
authors.forename: Michael G
authors.initials: MG
authors.lastname: Borland
authors.forename: Michael Eugene
authors.initials: ME
authors.lastname: Pugh
mesh_terms: D001671:Biochemistry / Q000193:education*; D001681:Biological Assay; D004247:DNA / Q000737:chemistry*; D004521:Educational Measurement; D018961:Educational Technology / Q000379:methods; D006801:Humans; D007753:Laboratories; D011506:Proteins / Q000737:chemistry*; D012313:RNA / Q000737:chemistry*; D012106:Research / Q000193:education / Q000295:instrumentation; D012996:Solutions / Q000737:chemistry; D013053:Spectrophotometry / Q000379:methods*; D013334:Students
chemical_list: D011506:Proteins; D012996:Solutions; D012313:RNA; D004247:DNA
keywords: Beer-Lambert- Bougert Law; Bial Assay; Bradford Assay; DNA and RNA determination; Dische Assay; protein determination