BIOL 2131 Study Guide - Final Guide: Lysis, Nucleic Acid Hybridization, Ethylenediaminetetraacetic Acid
16 May 2013
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Genomic dna- used for dna structure, dna hybridization, gentic studies and dna-protein interactions. Best way to extract dna (gold standard): marmur method: 2 barriers: cell membrane and cell wall, cell is disrupted from bacterial cell wall by physical or chemical methods (detergents and hydrolytic enzymes such as lysozymes) Gram negative cells (like e. coli) only require detergent: chlorofoam is used to remove protein components from the cell lysate. Second, it inhibits deoxyribonucleases that have a requirement for mg2+ or mn 2 1the cell pe. The separation between the phases can be seen through the sides of the plastic tube. Incubate for 20min at 37c (this is the temperature in which the enzyme functions) 20% sds (sodium dodecyl sulfate) is added to cell suspension incubate 10min at 50c in water bath this will break the lipid bilayer (polar and nonpolar ends) Sodium acetate- (5. 6 acidic substance) that will neutralize the ph.
