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DNA Composition Analysis

DNA Composition Analysis - me as I was able to see it...

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DNA Composition Analysis: G-C Content for Micrococcus lysodeikticus , Calf Thymus, and Salmon Testes Anna Krajec November 14, 2011 Lab Section 01
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Summary of Data Species % G+C % A+T T m (°C) Micrococcus lysodeikticus 71.48% 28.52% 89.99 °C Calf Thymus 49.80% 50.20% 80.32 °C Salmon Testes 46.70% 53.30% 79.05 °C
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Conclusions This lab made use of a newer technique to determine G-C content in DNA that does not require a thermal denaturation curve. We first denatured the DNA so that it would be single stranded. Then, using DNA nuclease, the heat-denatured DNA was digested into its 5’- mononucleotides. These nucleotides were separated on reverse phase HPLC columns. The technique for this lab was slightly over my head, but it was great to be able to get some exposure to new equipment. It is also helpful to see the way that development of new techniques can be so helpful in the laboratory. I still do not fully understand the way that the machine operated and how it is that the integrals of the curves give us the nucleotide content, but it became clearer to
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Unformatted text preview: me as I was able to see it hands-on. I learned a lot in this lab about G-C content and what it means for the strength of the DNA it is in. This was mostly done through observation of Tm and a thermal denaturation curve. We have learned in lecture that the 3 hydrogen bonds in G-C strengthen the DNA compared to the 2 hydrogen bonds between A and T. This, in turn, means that DNA with higher G-C content will require more heat to fully denature and thus have a higher Tm. I saw this after calculating the Xgc for each species and then plugging it into the Tm equation. Indeed, the one with the highest G-C content— Micrococcus lysodeikticus with 71.48%—had the highest Tm of 89.99 degrees. The one with the lowest—salmon testes with 46.70%--had the lowest Tm of 79.05 degrees. This lab just goes to show how big of an impact non-covalent, weak hydrogen bonds can have on a molecule....
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