What is the Difference Between Native and Denaturing Gel Electrophoresis

The key difference between native and denaturing gel electrophoresis is that in native gel electrophoresis, the biomolecule of interest is maintained its normal or native structure when running it through the gel, while in denaturing gel electrophoresis, the biomolecule of interest is not maintained its normal or native structure when running it through the gel.

Gel electrophoresis is a laboratory technique to separate DNA, RNA, or proteins according to their molecular size. In gel electrophoresis, the biomolecule of interest is pushed by an electrical field through a gel that contains small pores. Here, the shorter molecule moves faster than the longer molecule. This is because the shorter molecule migrates easily through the small pores in the gel. This property is called molecular sieving. Native and denaturing gel electrophoresis are two different types of gel electrophoresis techniques used to separate biomolecules such as DNA, RNA, and proteins.

CONTENTS

1. Overview and Key Difference
2. What is Native Gel Electrophoresis
3. What is Denaturing Gel Electrophoresis
4. Similarities – Native and Denaturing Gel Electrophoresis
5. Native vs Denaturing Gel Electrophoresis in Tabular Form
6. Summary – Native vs Denaturing Gel Electrophoresis

What is Native Gel Electrophoresis?

In native gel electrophoresis, the biomolecule (DNA, RNA, or protein) of interest maintains its normal or native structure when running through the gel. In native gel electrophoresis, the biomolecule is separated on the basis of both shape and length. Therefore, in this electrophoresis, biomolecules are separated based on the size, shape, and net charge of their native structure while preserving their function and activity.

Biomolecules like DNA, RNA, and proteins normally have a net negative charge. Biomolecules with a higher negative charge density will migrate faster. In addition, the smaller biomolecules will have a smaller frictional force as compared to larger biomolecules, so they, too, will migrate faster. As a result of this, biomolecules in native gel electrophoresis are separated based on both their mass and charge.

What is Denaturing Gel Electrophoresis?

In denaturing gel electrophoresis, the biomolecule of interest does not maintain its normal or native structure when running through the gel. It separates biomolecules on the basis of length. Denaturing gel electrophoresis destroys the complex structure of the biomolecules so that the molecules will separate only based on their mass when electrophoresed.

One common example of denaturing gel electrophoresis is SDS PAGE (SDS polyacrylamide gel electrophoresis), used for protein separation. In SDS PAGE, the protein samples are heated at 100^oC in the presence of anionic detergent such as SDS (sodium dodecyl sulfate). This reducing agent breaks the disulfide bonds of the proteins and destroys their quaternary protein structure. SDS also confers an overall negative charge to proteins. This process enables proteins to be separated based solely on their size or mass. SDS PAGE is useful to determine the molecular weight of the proteins as well.

What are the Similarities Between Native and Denaturing Gel Electrophoresis?

• Native and denaturing gel electrophoresis are two different types of gel electrophoresis techniques used to separate biomolecules such as DNA, RNA, and proteins.
• Both are molecular biological laboratory techniques.
• Both techniques should be performed by skilled technicians.
• The size of the biomolecules can be determined by using both gel electrophoresis techniques.
• Molecular ladders are used to determine the sizes of the biomolecules in both gel electrophoresis techniques.

What is the Difference Between Native and Denaturing Gel Electrophoresis?

In native gel electrophoresis, the biomolecule of interest maintains its normal or native structure when running through the gel, while in denaturing gel electrophoresis, the biomolecule of interest does not maintain its normal or native structure when running through the gel. Thus, this is the key difference between native and denaturing gel electrophoresis. Furthermore, reducing agents like SDS (sodium dodecyl sulfate) and DTT (dithiothreitol) are not used in native gel electrophoresis, while reducing agents like SDS (sodium dodecyl sulfate) and DTT (dithiothreitol) are used for denaturing in gel electrophoresis.

The below infographic presents the differences between native and denaturing gel electrophoresis in tabular form for side-by-side comparison.

Summary – Native vs Denaturing Gel Electrophoresis

Native and denaturing gel electrophoresis are two different types of gel electrophoresis techniques used to separate biomolecules such as nucleic acids and proteins. They are molecular biological laboratory techniques commonly used in modern laboratories. However, in native gel electrophoresis, the biomolecule of interest maintains its normal or native structure when running through the gel. In contrast, in denaturing gel electrophoresis, the biomolecule of interest does not maintain its normal or native structure when running through the gel. So, this is the key difference between native and denaturing gel electrophoresis.

Reference:

1. “Denaturing Gel Electrophoresis.” An Overview | ScienceDirect Topics.
2. “Native or Denaturing Gel – Which Is for You?” Advansta Inc.

Image Courtesy:

1. “Gel Electrophoresis” By Mckenzielower – Own work (CC BY-SA 4.0) via Commons Wikimedia
2. “SDS-PAGE Buffers” By Bensaccount at English Wikipedia (CC BY 3.0) via Commons Wikimedia