Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Restriction Enzymes

Restriction Enzymes

Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called restriction sites. Such palindromic DNA sequences or inverted repeats are commonly found in regions of functional significance, such as the origin of replication, gene operator sites, and regions containing transcription termination signals.
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Plasma nucleic acid analysis by massively parallel sequencing: pathological insights and diagnostic implications.

The Journal of pathology·2011

Same author

Introduction to the polymerase chain reaction.

Methods in molecular medicine·2011

Same author

Setting up a PCR laboratory.

Methods in molecular medicine·2011

Same author

Amplification from archival materials.

Methods in molecular medicine·2011

Same author

The amplification refractory mutation system.

Methods in molecular medicine·2011

Same author

Generation of labeled probes by PCR.

Methods in molecular medicine·2011

Same journal

Erratum to: Immunotherapeutic Approach to Cancer with Cutaneous DNA Vaccination.

Methods in molecular medicine·2015

Same journal

Methods for cancer gene therapy using tumor suppressor genes.

Methods in molecular medicine·2014

Same journal

Suppression of the human carcinoma phenotype by an antioncogene ribozyme.

Methods in molecular medicine·2014

Same journal

Methods for the use of stromal cells for therapeutic gene therapy.

Methods in molecular medicine·2014

Same journal

Methods for adenovirus-mediated gene transfer to synovium in vivo.

Methods in molecular medicine·2014

Same journal

Methods for gene transfer to synovium.

Methods in molecular medicine·2014

See all related articles

Search research articles

Related Experiment Video

Updated: Jun 3, 2026

A Robust Polymerase Chain Reaction-based Assay for Quantifying Cytosine-guanine-guanine Trinucleotide Repeats in Fragile X Mental Retardation-1 Gene

A Robust Polymerase Chain Reaction-based Assay for Quantifying Cytosine-guanine-guanine Trinucleotide Repeats in Fragile X Mental Retardation-1 Gene

Published on: September 16, 2019

Artificial Restriction Fragment Length Polymorphism (A-RFLP) Analysis.

Y M Lo¹, V A Horton

¹Department of Chemical Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR.

Methods in Molecular Medicine

|March 11, 2011

Summary

This summary is machine-generated.

Restriction fragment length polymorphism (RFLP) analysis is a classic method for analyzing DNA. Artificial RFLP (A-RFLP) creation using site-directed mutagenesis enables analysis of nearly all DNA polymorphisms.

More Related Videos

DNA Fingerprinting of Mycobacterium leprae Strains Using Variable Number Tandem Repeat (VNTR) - Fragment Length Analysis (FLA)

DNA Fingerprinting of Mycobacterium leprae Strains Using Variable Number Tandem Repeat (VNTR) - Fragment Length Analysis (FLA)

Published on: July 15, 2011

A Method to Study the C924T Polymorphism of the Thromboxane A2 Receptor Gene

A Method to Study the C924T Polymorphism of the Thromboxane A2 Receptor Gene

Published on: April 1, 2019

Related Experiment Videos

Last Updated: Jun 3, 2026

A Robust Polymerase Chain Reaction-based Assay for Quantifying Cytosine-guanine-guanine Trinucleotide Repeats in Fragile X Mental Retardation-1 Gene

A Robust Polymerase Chain Reaction-based Assay for Quantifying Cytosine-guanine-guanine Trinucleotide Repeats in Fragile X Mental Retardation-1 Gene

Published on: September 16, 2019

DNA Fingerprinting of Mycobacterium leprae Strains Using Variable Number Tandem Repeat (VNTR) - Fragment Length Analysis (FLA)

DNA Fingerprinting of Mycobacterium leprae Strains Using Variable Number Tandem Repeat (VNTR) - Fragment Length Analysis (FLA)

Published on: July 15, 2011

A Method to Study the C924T Polymorphism of the Thromboxane A2 Receptor Gene

A Method to Study the C924T Polymorphism of the Thromboxane A2 Receptor Gene

Published on: April 1, 2019

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

Restriction analysis of polymerase chain reaction (PCR) products is an established technique for analyzing amplification products.
This method is effective for distinguishing alleles when polymorphisms alter restriction enzyme sites.
A significant limitation is that many polymorphisms do not affect restriction sites, rendering them unsuitable for standard analysis.

Purpose of the Study:

To present a method for analyzing DNA polymorphisms that are not amenable to traditional restriction enzyme analysis.
To introduce the concept and principles of creating artificial restriction fragment length polymorphisms (A-RFLP).

Main Methods:

Utilizing site-directed mutagenesis with primers containing mismatches near the 3' ends of PCR products.
This technique aims to introduce or abolish restriction enzyme sites artificially.
The process generates an artificial restriction fragment length polymorphism (A-RFLP) for nearly all naturally occurring DNA polymorphisms.

Main Results:

The described method allows for the creation of A-RFLP for a wide range of DNA polymorphisms.
This approach overcomes the limitations of standard RFLP analysis where polymorphisms do not alter restriction enzyme sites.
The principles illustrated demonstrate the feasibility of generating detectable fragment length differences.

Conclusions:

Artificial RFLP (A-RFLP) provides a versatile strategy to analyze DNA polymorphisms previously undetectable by standard restriction analysis.
This technique significantly expands the applicability of RFLP-based genotyping and genetic analysis.
The method offers a powerful tool for molecular biology research and diagnostics.