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

Search research articles

Related Experiment Videos

Enzymatic mutation detection in the P53 gene.

M Inganäs¹, S Byding, A Eckersten

¹Amersham Pharmacia Biotech, SE-751 84 Uppsala, Sweden.

Clinical Chemistry

|October 6, 2000

Summary

The enzymatic mutation detection (EMD) assay accurately identifies genetic variations in DNA fragments over 1kb. This method offers high sensitivity and specificity for detecting mutations in genes, regardless of their location or type.

Related Concept Videos

You might also read

Related Articles

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

Sort by

Same author

High risk of permanent stoma after anastomotic leakage in anterior resection for rectal cancer.

Colorectal disease : the official journal of the Association of Coloproctology of Great Britain and Ireland·2018

Same author

Late leakage after anterior resection: a defunctioning stoma alters the clinical course of anastomotic leakage.

Colorectal disease : the official journal of the Association of Coloproctology of Great Britain and Ireland·2017

Same author

The importance of rectal washout for the oncological outcome after Hartmann's procedure for rectal cancer: analysis of population-based data from the Swedish Colorectal Cancer Registry.

Techniques in coloproctology·2017

Same author

Time trends, improvements and national auditing of rectal cancer management over an 18-year period.

Colorectal disease : the official journal of the Association of Coloproctology of Great Britain and Ireland·2015

Same author

Lymph node tissue kallikrein-related peptidase 6 mRNA: a progression marker for colorectal cancer.

British journal of cancer·2012

Same author

A Longitudinal Study of Perceived Health during Pregnancy: Antecedents and Outcomes.

Journal of health psychology·2011

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

The enzymatic mutation detection (EMD) assay utilizes T4 endonuclease VII to cleave DNA heteroduplexes at mismatch sites.
This enzymatic cleavage generates smaller fragments, enabling detection via gel electrophoresis.
EMD can identify single/multiple base changes, insertions, and deletions.

Purpose of the Study:

To evaluate the sensitivity, specificity, and positional accuracy of the EMD assay.
To assess EMD performance across different analytical platforms.
To determine the efficacy of EMD for detecting mutations in long PCR products.

Main Methods:

EMD assay with the PASSPORT(TM) Mutation Scanning Kit was performed on PCR products from P53 and IL4r genes.

Related Experiment Videos

Analysis was conducted on radioactive detection, ALFexpress, and ABI PRISM 377 platforms.

Results were compared against cDNA sequencing data.

Main Results:

EMD demonstrated >90% sensitivity and >80% specificity across all platforms.
Mutations were localized to within two codons (automated sequencers) or four codons (radioactive detection).
The assay successfully detected at least five mismatches in heteroduplexes larger than 1 kb.

Conclusions:

The EMD system efficiently detects genetic variations in DNA fragments >1 kb, irrespective of location or type.
This technology is highly suitable for detecting mutations in genes with unpredictable mutation sites.
EMD provides a robust method for comprehensive genetic variation analysis.