Electrophoresis Right Ans - A separation technique that is based on the
movement of charged particles in an electric field.
Electromigration Right Ans - At any given pH, charged molecules may exist
as anions (move to anode) or cations (move to cathode)
Electromotive Force Right Ans - Basis of Electrophoresis. Force that pushes
molecule through the matrix based on their charge.
Factors affecting rate of migration of the ions Right Ans - Net charge
Size and shape
Buffer pH
Strength of electric field
Properties of medium
Temperature of system
Types of Buffers Right Ans - Barbitone (serum protein)
Phosphate (enzyme)
TBE + TAE (nucleic acids)
Tris-glycine (proteins)
Media Right Ans - Paper
Agar
Cellulose acetate strip
Starch
Agarose
Acrylamide
Types of Electrophoresis Right Ans - Moving boundary/frontal
Zonal (ones we use)
Immuno Electrophoresis
Moving Boundary/ Frontal Electrophoresis Right Ans - Solution injected in
U tube
Arms filled with buffer
, When electricity passed, separate into bands of proteins, no discrete zones.
Zonal Electrophoresis Right Ans - Sample as narrow band
Separation in form of discrete bands
Support media.
Paper Electrophoresis Right Ans - Volume placed along line drawn on
Whatmann paper.
Ends soaked in buffer.
Electricity passed.
Starch Gel Electrophoresis Right Ans - Isoenzymes.
Partially hydrolysed potato starch is used.
Opaque
Non denatured proteins can be separated.
Visualised via staining.
Cellulose Acetate Electrophoresis Right Ans - Strips of cellulose acetate.
Better resolving power
No absorption
No trailing, transparent
Used for plasma proteins.
Gel Electrophoresis Right Ans - Gelatinous material such as agarose,
acrylamide or cellulose acetate.
Support medium to separate samples.
Agarose Gel Electrophoresis Right Ans - Analyse DNA fragments.
Gel from agarose (derivative of agar)
Pores act as a seive and affect the mobility of sample on the medium. Form
bands. No electric charge.
Polyacrylamide Gel Electrophoresis (PAGE) Right Ans - Acrylamide and a
cross linker, methylene bis-acrylamide. Synthetic. Resolves 1bp difference.
Capillary Gel Electrophoresis Right Ans - Thin glass capillary (25 to 100
micrometer diameter)
Polyacrylamide for sieving
Rapid than slab gels