Gel Electrophoresis

Question 1

What are the 4 methods used to analyse and separate soluble components (proteins and nucleic acids) using electrophoretic techniques?

Answer 1

Agarose gel electrophoresis
Polyacrylamide gel electrophoresis
Isoelectric focusing
Capillary electrophoresis

Question 2

What is electrophoresis?

Answer 2

• A technique used to separate DNA
  fragments (or other macromolecules,
  such as RNA and proteins) based on
  their size and charge
• We can separate molecules within a
  gel or fluid by using an electric field.
• The rate and direction of the particles
  moving within the gel will be
  dependent on their size and
  molecular charge
  it uses 2 electrodes- cathode (-) and anode(+) and an electrolyte gel which serves as a conducting medium

Question 3

What can be separated?

Answer 3

Amino acids- Peptides and Proteins
Nucleotides- DNA and RNA
Nanoparticles

Question 4

What is the difference between peptides and proteins?

Answer 4

Proteins are more than 30 base pairs whereas peptides contain less than 30 base pairs

Question 5

Discuss the methods of separation of the electrophoretic techniques

Answer 5

AGE- size and charge
PAGE - size and charge
IEF- pH and pI (isoelectric point)
CE - size and charge

Question 6

Discuss the different conducting mediums used by electrophoretic techniques

Answer 6

AGE-agarose gel
PAGE- polyacrylamide gel
IEF electricity
CE- pH solution

Question 7

What is the product of different electrophoretic techniques?

Answer 7

AGE- DNA
PAGE- protein
IEF -protein isoforms