bme100 - BME 100L. Modeling Cellular and Molecular Systems...

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BME 100L.  Modeling Cellular and Molecular Systems Spring 2007 Lab 1. Dissociation of Double Stranded Polynucleotides The goal of this laboratory is to measure the dissociation of polyinosinic acid- polycytidinic acid (polyI-polyC) as a function of temperature. Inosinic acid is a derivative of adenosine and can form hydrogen bonds with the nucleotides cytidine, uridine, and adenosine. The structure of these molecules is shown below. O O P OH OH O OH OH N N N NH Inosinic acid Cytidinic acid O O P H N 2 Hydrogen bonds Dissociation can be measured by taking advantage of the hypochromicity of the polynucleotides (i.e., decrease light absorption by the solution) upon double helix formation. A. Prelab 1. Review laboratory procedure. 2. Identify any questions that you may have for the TAs. 3. Form a lab team with another student. B. Experiment 1. The following buffer is available: 10 mM Tris 10 mM EDTA 20 mM NaCl Adjust pH to 6.8 2. A stock solution of 0.762 mM polyinosinic acid-polycytidinic acid (polyI-polyC) was prepared in the buffer. The formula weight of polyI-polyC is 671.41 g (mole base pair)
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bme100 - BME 100L. Modeling Cellular and Molecular Systems...

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