Biochemical methods Flashcards
What are biochemical methods used for?
To study the structure, function, and interactions of biological molecules such as proteins, nucleic acids, and metabolites.
What are the key principles of biochemical methods?
Specificity: Target-specific detection or separation.
Sensitivity: Ability to detect low concentrations.
Reproducibility: Consistent results across experiments.
What are the common types of biochemical methods?
Chromatography (e.g., HPLC, gas chromatography).
Electrophoresis (e.g., SDS-PAGE, agarose gel electrophoresis).
Spectroscopy (e.g., UV-Vis, fluorescence, NMR).
What is chromatography?
A separation technique based on the differential partitioning of molecules between a stationary phase and a mobile phase.
What are the types of chromatography?
Ion-exchange chromatography: Separates based on charge.
Size-exclusion chromatography: Separates based on size.
Affinity chromatography: Separates based on specific interactions (e.g., antibody-antigen).
What are the key applications of chromatography?
Protein purification.
Metabolite analysis.
Identification of small molecules.
What is spectroscopy?
A method that uses the interaction of light with matter to analyse molecular properties
What are the types of spectroscopy used in biochemistry?
UV-Vis spectroscopy: Measures absorbance at specific wavelengths.
Fluorescence spectroscopy: Detects emitted light from excited molecules.
NMR spectroscopy: Analyzes molecular structure using magnetic fields.
What is electrophoresis?
and common types
A technique used to separate charged molecules under an electric field.
SDS-PAGE: Separates proteins based on size.
Agarose gel electrophoresis: Separates nucleic acids based on size.
How is chromatography used in labs?
Protein purification using affinity tags.
Metabolite profiling using HPLC.
What are the practical challenges in electrophoresis?
Smearing due to overloaded samples.
Inconsistent band resolution from uneven gel casting.
What is mass spectrometry?
A technique for determining the mass-to-charge ratio of ions to analyze molecular structure and composition.
What are the applications of mass spectrometry?
Identifying post-translational modifications.
Quantifying metabolites and proteins.
