Separation techniques Centrifugation DIFFERENTIAL CENTRIFUGATION DENSITY GRADIENT CENTRIFUGATION

  • Slides: 13
Download presentation
Separation techniques

Separation techniques

Centrifugation • DIFFERENTIAL CENTRIFUGATION • DENSITY GRADIENT CENTRIFUGATION

Centrifugation • DIFFERENTIAL CENTRIFUGATION • DENSITY GRADIENT CENTRIFUGATION

DIFFERENTIAL CENTRIFUGATION

DIFFERENTIAL CENTRIFUGATION

DENSITY GRADIENT CENTRIFUGATION

DENSITY GRADIENT CENTRIFUGATION

Meselson-Stahl Experiment

Meselson-Stahl Experiment

Gel Electrophoresis

Gel Electrophoresis

Definition • Electro = Charge + Phorsesis= Carry • Electrophoresis = Separation of charged

Definition • Electro = Charge + Phorsesis= Carry • Electrophoresis = Separation of charged molecules by differences in their rate of migration in an electric field.

The Components of The System • Molecules to be separated • Proteins • Nucleic

The Components of The System • Molecules to be separated • Proteins • Nucleic Acids • Support medium • Gel (Starch, Polyacrylamide or Agarose) • Buffer System • DC Power Source

Common Support Media for Biological Molecules • Proteins • Native Gel (Acrylamide or Starch)

Common Support Media for Biological Molecules • Proteins • Native Gel (Acrylamide or Starch) • Denaturing (SDS) Gel (Acrylamide) • Nucleic Acids – DNA & RNA • Agarose Gel • Acrylamide Gel

Factors that Influence Mobility • Properties of the Molecules to be separated • Molecular

Factors that Influence Mobility • Properties of the Molecules to be separated • Molecular size (MW) • Molecular shape • Molecular charge • Properties of the System • Electric field strength (V/cm) • Porosity of the support medium (% S) • Conductivity of the buffer (R)

These Factors Interact • Mobility is proportional to charge/MW. • Mobility is proportional to

These Factors Interact • Mobility is proportional to charge/MW. • Mobility is proportional to field strength.

SDS Polyacrylamide Gel Electrophoresis (SDS PAGE)

SDS Polyacrylamide Gel Electrophoresis (SDS PAGE)

Separation of DNA Molecules in Agarose Gels • The phosphate groups bear negative charge

Separation of DNA Molecules in Agarose Gels • The phosphate groups bear negative charge at neutral p. H (2 phosphates/BP) • Therefore mobility will be based on number of base pairs/molecule

Centrifugation Centrifugation involves separation of liquids and particlesCentrifugation Centrifugation involves separation of liquids and particles
Centrifugation Centrifugation involves separation of liquids and particlesCentrifugation Centrifugation involves separation of liquids and particles
Chemistry SEPARATION TECHNIQUES Separation Techniques Involve the separationChemistry SEPARATION TECHNIQUES Separation Techniques Involve the separation
Separation Process Separation Methods Separation Methods 1 SeparationSeparation Process Separation Methods Separation Methods 1 Separation
SEPERATION OF BIOMOLECULES Centrifugation Sedimentation density gradient ChromatographySEPERATION OF BIOMOLECULES Centrifugation Sedimentation density gradient Chromatography
DENSITY Density and Buoyancy Key Points Density DensityDENSITY Density and Buoyancy Key Points Density Density
Gradient Methods Steepest Descent gradient descent Conjugate GradientGradient Methods Steepest Descent gradient descent Conjugate Gradient
CENTRIFUGATION Technique de centrifugation Maria AGUILA Anas AYMONTCENTRIFUGATION Technique de centrifugation Maria AGUILA Anas AYMONT
CENTRIFUGATION Dr Vinod K V Types of centrifugationCENTRIFUGATION Dr Vinod K V Types of centrifugation
Separation Zone Inshore Traffic Zone Separation Line SeparationSeparation Zone Inshore Traffic Zone Separation Line Separation
Vocabulary Density Gradient The change in density inVocabulary Density Gradient The change in density in
Partial Differential Total Differential Total Differential of FunctionPartial Differential Total Differential Total Differential of Function