GEL ELECTROPHORESIS

Biotechnology
GEL ELECTROPHORESIS CAN BE USED
FOR
WHAT THE GEL TRAY IS MADE OF
 WHAT IS GEL ELECTROPHORESIS?

• Electrophoresis is a technique in biotechnology

• used to separate & sometimes purify macromolecules
• Macromolecules such proteins and nucleic acids
• Macromolecules that differ in size, charge or conformation.
 PURPOSE

• A method used for the separation and isolation of DNA

fragments
• Can be used to separate the size of
1. DNA
2. RNA
3. Protein
• We will be using it to purify DNA, RNA AND PROTEINS.
TECHNIQUE
 DNA LADDER

• A molecular-weight size
marker, also referred to as
a protein ladder, DNA ladder,
or RNA ladder, is a set
of standards that are used to
identify the approximate size of
a molecule run on
a gel during electrophoresis
• The principle - molecular weight is
inversely proportional to
migration rate through a gel
matrix.
 2 MOST COMMON GEL TYPES
AGAROSE
• Used for separating big pieces of DNA
• Usually for DNA fragments that are
bigger than 50bp
• Poured Horizontally
• Separate large molecules
• Non-toxic
• Mostly used for DNA separation
• Staining step: before or after pouring
the gel
• Ethidium bromide is mostly used
SDS PAGE
• sodium dodecyl sulfate polyacrylamide gel
• Used for separating small pieces of DNA and proteins
• sodium dodecyl sulfate – reagent that denature protein > disrupting
any non covalent interactions they have >> this doesn’t matter for
whatever charge they may have but will be separated based on their
size
• PAGE > polyacrylamide gel electrophoresis
• Poured Vertically
• Separate small molecules
• Neurotoxin
• Used for DNA or Protein separation
• Staining step: after pouring the gel
• Coomassie blue stain is mostly used
WHY IS IT IMPORTANT TO REMOVE ALL THE
CONTENTS OF YOUR TUBES BEFORE
ATTEMPTING TO LOAD A GEL?
HOW COULD GEL ELECTROPHORESIS BE
APPLIED TO THE IDENTIFIC ATION OF A
GENE MUTATION?
WHAT ALLOWS A CURRENT TO TRAVEL
THROUGH THE GEL TRAY