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# A mechanical method of seperating immiscible
liquids or solids from liquids,by the application of centrifugal force.
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# It is a piece of equipment,generally
driven by an electric motor,that puts an object in rotation around a fixed axis, applying a force perpendicular to the axis.
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# Isolating suspensions,
# Isotope seperation, # Aeronautics & astronautics, # Earthquake & blast stimulation & # Commercial applications.
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# This technique concerned with the seperation,isolation &
purification of subcellular organelles,whole plasma membrane,polysomes,ribosomes,chromatin,nucleic acids, lipoproteins & viruses.
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# This technique concerned with the study of pure, or
virtually pure macromolecules or particles.
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ROTORS: # Swinging – bucket rotors, # Fixed – angle rotors,&
# Vertical rotors. ROTORS:
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SWINGING – BUCKET ROTORS:
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FIXED – ANGLE ROTORS:
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VERTICAL ROTORS:
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VERTICAL ROTORS:
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# Density gradient centrifugation,
# Rate-zonal seperation, # Isopycnic seperation. # Differential centrifugation, # K – factor centrifugation, # Sucrose gradient centrifugation, # Microcentrifugation, # Superspeed centrifugation,& # ultracentrifugation.
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# It is useful to purify subcellular organelles & macromolecules.
# It is also useful for the determinaton of buoyant densities,& # It is also useful for the estimation of the sedimentation coefficient.
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1) Seperation of : # Proteins, # Enzymes, # Hormones, # RNA – DNA hybrids, # Ribosomal subunits,& # Subcellular organelles. 2)For the analysis of size distribution of samples of polysomes & lipoprotein fractionations.
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# To determine the buoyant density of a particle,
# To determine the base composition of the double – stranded DNA, # To separate & purify viruses, # To analyze nucleic acids.
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centrifugation Sample + Caesium chloride Cscl molecules sedimentation Rise or sediment Concentration gradient (density gradient) Solution density = Own buoyant density
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It is the most commonly used method,for the isolation of
subcellular organelles from the homogenised tissue.
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# It can be used to determine the time required for pelleting a
fraction,with a known sedimentation coefficient.
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# It is used to purify enveloped viruses & ribosomes.
# It will also separate cell organelles from crude cellular extracts. # It will also purify exosomes.
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# These centrifuges are run in batch to isolate small volumes of
biological macromolecules or cells. # It will purify cells & nuclei.
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# It can be used to isolate subcellular organelles such as
endoplasmic reticulum,ribosomes,cytoplasmic components, smaller particles such as proteins & to harvest virus in pure form.
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# Theodor Svedberg(1925) - Analytical centrifuge
# Edward Greydon Pickles - Vacuum centrifuge Pickles(1946) Spinco(1946) Model L Beckman instruments (1954)
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# It is used for the determination of the macromolecular including
amino acid composition of protein,protein’s current conformation, & size distribution of macromolecular samples.
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They are used in biology for pelleting of fine particulate fractions.
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# It is the most important widely applied research techniques in
biochemistry,cellular & molecular biology, & in medicine.
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