Sign In

Molecular Cloning

Overview

Molecular cloning is a set of methods, which are used to insert recombinant DNA into a vector - a carrier of DNA molecules that will replicate recombinant DNA fragments in host organisms. The DNA fragment, which may be a gene, can be isolated from a prokaryotic or eukaryotic specimen. Following isolation of the fragment of interest, or insert, both the vector and insert must be cut with restriction enzymes and purified. The purified pieces are joined together though a technique called ligation. The enzyme that catalyzes the ligation reaction is known as ligase.

This video explains the major methods that are combined, in tandem, to comprise the overall molecular cloning procedure. Critical aspects of molecular cloning are discussed, such as the need for molecular cloning strategy and how to keep track of transformed bacterial colonies. Verification steps, such as checking purified plasmid for the presence of insert with restrictions digests and sequencing are also mentioned.

Procedure

Molecular cloning is a set of techniques used to insert recombinant DNA from a prokaryotic or eukaryotic source into a replicating vehicle such as plasmids or viral vectors. Cloning refers to making numerous copies of a DNA fragment of interest, such as a gene. In this video you will learn about the different steps of molecular cloning, how to set up the procedure, and different applications of this technique.

At least two important DNA molecules are required before cloning begins. First,

Log in or to access full content. Learn more about your institution’s access to JoVE content here

Tags
Molecular CloningRecombinant DNAProkaryoticEukaryoticReplicating VehiclePlasmidsViral VectorsGene CloningDNA FragmentInsertFunction Of A GeneVectorMultiple Cloning SiteRestriction EndonucleasesLigationOrigin Of ReplicationAntibiotic Gene

Skip to...

0:00

Overview

0:42

Basic Principles

3:47

Procedure

7:45

Applications

9:34

Summary

Videos from this collection:

article

Now Playing

Molecular Cloning

Basic Methods in Cellular and Molecular Biology

372.1K Views

article

Using a Hemacytometer to Count Cells

Basic Methods in Cellular and Molecular Biology

216.4K Views

article

Passaging Cells

Basic Methods in Cellular and Molecular Biology

185.4K Views

article

PCR: The Polymerase Chain Reaction

Basic Methods in Cellular and Molecular Biology

701.0K Views

article

DNA Gel Electrophoresis

Basic Methods in Cellular and Molecular Biology

589.6K Views

article

Separating Protein with SDS-PAGE

Basic Methods in Cellular and Molecular Biology

466.0K Views

article

Bacterial Transformation: The Heat Shock Method

Basic Methods in Cellular and Molecular Biology

728.2K Views

article

Bacterial Transformation: Electroporation

Basic Methods in Cellular and Molecular Biology

113.8K Views

article

The ELISA Method

Basic Methods in Cellular and Molecular Biology

247.6K Views

article

Plasmid Purification

Basic Methods in Cellular and Molecular Biology

300.6K Views

article

Gel Purification

Basic Methods in Cellular and Molecular Biology

108.0K Views

article

The Western Blot

Basic Methods in Cellular and Molecular Biology

495.3K Views

article

An Introduction to Transfection

Basic Methods in Cellular and Molecular Biology

167.8K Views

article

DNA Ligation Reactions

Basic Methods in Cellular and Molecular Biology

185.0K Views

article

Restriction Enzyme Digests

Basic Methods in Cellular and Molecular Biology

277.5K Views

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2025 MyJoVE Corporation. All rights reserved