Electrophoresis Flashcards
Electrophoresis
Movement of charged particles through the porous gel matric under the influence of electric field
Name the two types of electrophoresis
Moving boundary electrophoresis
Zone electrophoresis
What is moving boundary electrophoresis?
The electrophoresis that doesn’t need any supporting media and forms boundary
What is zone electrophoresis
The electrophoresis that needs a supporting media and forms bands
What are the two types of gel matrix?
Polyacrylamide
Agarose
What is the correlation between concentration and the molecular weight of the sample?
Inversely proportional
How is polyacrylamide formed?
Acrylamide + bisacrylamide
Name the initiator and a catalyst in the formation of polyacrylamide chain.
Initiator - ammonium persulfate
Catalyst - N, N, N’, N’ - Tetramethylene diamine
What is the concentration used in separation of protein in polyacrylamide gel?
10 to 15%
Is angrose gel linear or branched polysaccharide?
Linear
How is agarose gel formed?
Galactose and anhydrogalactose
Name the algae used in the formation of agarose.
Gracilaria and Gelidium
What is the concentration of agarose gel?
1 to 3%
What are the stains used in agarose gel electrophoresis?
Ethidium bromide, DAPI, methylene blue
Name the intercalating agent (or metal chelater) of agarose gel electrophoresis
Ethidium bromide
What kind of detergent is sodium dodecyl sulphate?
Anionic detergent
Name these tracking dye in SDS electrophoresis
Bromo phenol blue
Write the movement order in SDS electrophoresis
Cl — Protein SDS — Glycinate
Name the anionic dye that binds to the hydrophobic part of the protein
Coomasie brilliant blue
Name the dye for the sensitive proteins of ng
Silver stain
Name two reducing agents
B mercaptoethanol and dithiothretol
In isoelectric focusing who create the pH gradient?
Polyampholyte
Write the relation between the charge and isoelectric point of a protein
pI more than pH then positive
pI less than pH then negative
pI equals to pH then neutral
In 2D electrophoresis how are the proteins separated?
First by pI and the by mass
Why are DNA sequencing gel used?
To seperate single stranded DNA that are shorter than 1000nt and differing in size by 1nt
Why is pulsed field electrophoresis used?
To separate molecules that are of size 2000kbp to 10Mbp
What is the unique characteristic of pulsed field gel electrophoresis?
Uses electric field at an angle of 120°
What can we use when the sample is sensitive to the ammonium persulfate?
Riboflavin
Who is a neurotoxin in their monomer form?
Bisacrylamide and acrylamide
Blue native PAGE
The technique that allows separation of proteins present in multiprotein complexes in a native confirmation with higher resolution
Name the two buffer for the separation of nucleic acid
TBE and TAE
Name the salt that creates favourable ionic strength and pH for the sample
Tris HCl
Name the density ingredients of the gel
Glycerol or sucrose
How is capillary electrophoresis of DNA is better than the normal electrophoresis?
Provides better speed, resolution, sensitivity and data handling
