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Exam_2_s05 - Spring'05 CH370 CH387D Name Hackert Exam II...

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Spring '05 CH370 / CH387D Name _____________________ Hackert Exam II UTID ________________ R = 8.314 x 10 7 g-cm 2 /(sec 2 -mol-K); RCF = (1.119 x 10 -5 )(rpm) 2 (r); η = 0.01 g/(cm-sec) ρ water = 1.00 g/cm 3 ; s = M(1 - ν ρ )/ N o f ; (1/c r )(dc r /dr) = M ω 2 r(1 - ν ρ )/RT; k = 1.38 x 10 -23 J/K; h = 6.63 x 10 -34 J-sec; A = A o exp(-kt); k = ln(2)/ τ ½ 1. A solution has a %T of 42%. What is the absorbance? (3) An extinction coefficient is given as an E 0.1% extinction coefficient for λ = 440 nm with a magnitude of 3.55. Calculate the concentration in mg/mL of a solution that measures an absorbance of 0.50 at 440 nm. (5) 2. Briefly explain the advantages of using an expression vector that incorporates an added “His 6 -tag” at the N-terminus of your desired protein. Include in your explanation the type of column you would use and the methods required for eluting your protein during the purification procedures. (5) 3. In what order would the following proteins be eluted from a DiEthylAminoEthyl (DEAE)-Sephadex column using first a buffer wash at pH 7.2 followed by an increasing salt gradient? Protein: Lactate DH Cytochrome c Tyrpsin pI : 10.5 7.2 2.5 (4) (Off first) ______________ ________________ ______________ (Off Last)
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4. Consider a FRET experiment where the measured efficiency of energy transfer between chromophores BevoOrange and AggieMaroon is only 8%. If Ro = 26.7Å for these two chromophores, calculate the expected separation between the Orange and Maroon.
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