11a_Lab%203

11a_Lab 3 - Lab 3 Dissociation of Double-Stranded Polynucleotides A Thermodynamic Lab S1 = polyinosinic acid(polyI a derivative of adenosine S2 =

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1 Lab 3: Dissociation of Double-Stranded Polynucleotides A Thermodynamic Lab Initially, [D] = C 0 (molar concentration), [S 1 ] = [S 2 ] = 0 During the reaction, the total polynucleotide concentration is C ts = 2[D] + [S 1 ] + [S 2 ] = 2[D] + [S] = 2C 0 S 1 = polyinosinic acid ( polyI ), a derivative of adenosine S 2 = polycytidinic acid ( polyC ) ts D C ] D [ f 2 (fraction of polynuleotides in double stranded form) How to measure f D or [D]? DS 1 + S 2 UV light at 251 nm Measure the intensity I of light using the spectrophotometer. I 0 Absorbance I I log A 0 10 (reported by the spectrophotometer) Beer’s law cuvette h is the cuvette thickness, is the extinction coefficients, c is the concentration of solute. In a mixture of D and S ) ] S [ ] D [ ( h A A S D b 251 251 2 or   S D D D ts b ) f ( f hC A A 251 251 1 ) c h exp( I ) kh exp( I I j j j 0 0 j j j c h A j j j b c h A (without D and S)
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2 ) /( hC A A f S D S ts b D 251 251 251 Thus, where A and A b can be measured. C ts and h are known in the preparation of experiment.
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This note was uploaded on 01/17/2010 for the course BME 100 taught by Professor Yuan during the Fall '07 term at Duke.

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11a_Lab 3 - Lab 3 Dissociation of Double-Stranded Polynucleotides A Thermodynamic Lab S1 = polyinosinic acid(polyI a derivative of adenosine S2 =

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