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19 EXERCISE 2 DETERMINATION OF PROTEIN CONCENTRATION “What can I wish for the youth of my country who devote themselves to science? Firstly, gradualness. About this most important condition of fruitful scientific work I never can speak without emotion. Gradualness, gradualness, gradualness. From the very beginning of your work, school yourselves to severe gradualness in the accumulation of knowledge. Learn the ABC of science before you try to ascend to its summit. Learn, compare, collect the facts! Secondly, modesty. Never think that you already know all. However highly you are appraised, always have the courage to say to yourself - I am ignorant. Thirdly, passion. Remember that science claims a man's whole life. Had he two lives they would not suffice. Science demands an undivided allegiance from its followers. In your work and in your research there must always be passion.” Ivan Petrovich Pavlov (1849-1936) * OPERATION OF THE SPECTROPHOTOMETER * PREPARATION OF A STANDARD CURVE, ESTIMATING UNKNOWNS * THE MCIROLITER PIPETTE * PHENOTYPE OF F1 SEEDILINGS Equipment and Supplies Protein Concentration spectrophotometer Bradford dye color wheel protein solution (BSA 200μg/ml) water in squirt bottles parafilm, scissors and rulers beakers test tubes and racks 10 ml pipettes and pipettors unknown protein solutions Vortex mixer Microliter pipettes P20, P200, P1000 Phenotype of F1 Plants Parental and F1s The purpose of this exercise is to measure the concentration of protein in solution , to digest DNA from bacteriophage lambda with a restriction enzyme, and to pollinate the F1 flowers to produce the F2 generation.
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Exercise 2. Protein Concentration 20 PART I. Determination of Protein Concentration All living organisms contain proteins which are composed of amino acids joined together by peptide bonds . Without proteins, life as we know it is not possible. Proteins function as: structural components of cells and tissue, components of motility and contraction, chemical messengers (hormones), receptors of messages on the cell surface or interior, transporters of other substances, catalysts which carry out many of the chemical reactions required to maintain life, defense molecules (antibodies) regulators of cellular processes. In order to understand how living organisms operate, a complete knowledge of all the thousands of proteins which cells make and use is required. This is a formidable task. Many research scientists worldwide are presently engaged in identifying, purifying and characterizing proteins. In your lifetime, enormous progress will be made in our understanding of proteins and some of you will contribute to this effort. Regardless of what protein is under investigation, it is very important for a biochemist to be able to determine the concentration of proteins in a solution. One of the most commonly used methods was developed in 1976
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This note was uploaded on 09/28/2010 for the course SCI mcdb 1a & taught by Professor Bush during the Spring '10 term at UCSB.

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