Describe how you would prepare 500mL Edward’s buffer when you have 1M Tris-CL (pH 7.5), 1M NaCl, 50mM EDTA, and 10% SDS in the lab. the final concentration in Edward’s buffer: 200mM Tris-Cl, pH 7.5: lysis buffer, regulates pH and osmolality 250mM NaCl: removes protein bound to DNA 25mM EDTA (Ethylenediaminetetraacetic acid): Chelating agent that depletes metal ions 0.5% SDS (sodium dodecyl sulfate): surfactant commonly found in cleaning products. In this lab it is used to lyse cells.
Enzyme kinetics
In biochemistry, enzymes are proteins that act as biological catalysts. Catalysis is the addition of a catalyst to a chemical reaction to speed up the pace of the reaction. Catalysis can be categorized as either homogeneous or heterogeneous, depending on whether the catalysts are distributed in the same phase as that of the reactants. Enzymes are an essential part of the cell because, without them, many organic processes would slow down and thus will affect the processes that are important for cell survival and sustenance.
Regulation of Enzymes
A substance that acts as a catalyst to regulate the reaction rate in the living organism's metabolic pathways without itself getting altered is an enzyme. Most of the biological reactions and metabolic pathways in the living systems are carried out by enzymes. They are specific for their works and work in particular conditions. It maintains the best possible rate of reaction in the most stable state. The enzymes have distinct properties as they can proceed with the reaction in any direction, their particular binding sites, pH specificity, temperature specificity required in very few amounts.
Describe how you would prepare 500mL Edward’s buffer when you have 1M Tris-CL (pH 7.5), 1M NaCl, 50mM EDTA, and 10% SDS in the lab.
- the final concentration in Edward’s buffer:
- 200mM Tris-Cl, pH 7.5: lysis buffer, regulates pH and osmolality
- 250mM NaCl: removes protein bound to DNA
- 25mM EDTA (Ethylenediaminetetraacetic acid): Chelating agent that depletes metal ions
- 0.5% SDS (sodium dodecyl sulfate): surfactant commonly found in cleaning products. In this lab it is used to lyse cells.
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