Mutation analysis of genetic diseases by asymmetric-PCR SSCP and ethidium bromide staining

application to neurofibromatosis and cystic fibrosis

Conxi Lázaro, Xavier P. Estivill

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The Single Strand Conformation Polymorphism (SSCP) technique is widely used in mutation analysis. We have introduced several modifications to the SSCP method, which overcome the problem of incomplete denaturation or reannealing of DNA during electrophoresis. The modifications consist of asymmetrical PCR amplification of the sequence of interest, electrophoresis with a higher concentration of acrylamide, and the analysis of the DNA fragments under u.v. light. We have applied this method to the analysis of two specific diseases: neurofibromatosis type 1 (NF1) and cystic fibrosis (CF) from PCR amplified exons. Two single nucleotide changes were observed with this method.

Original languageEnglish
Pages (from-to)357-359
Number of pages3
JournalMolecular and Cellular Probes
Volume6
Issue number5
DOIs
Publication statusPublished - 1992
Externally publishedYes

Fingerprint

Inborn Genetic Diseases
Neurofibromatoses
Ethidium
Cystic Fibrosis
Staining and Labeling
Polymerase Chain Reaction
Mutation
Electrophoresis
Neurofibromatosis 1
Acrylamide
DNA
Exons
Nucleotides
Light

Keywords

  • cystic fibrosis
  • mutation analysis
  • neurofibromatosis type 1
  • SSCP

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

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abstract = "The Single Strand Conformation Polymorphism (SSCP) technique is widely used in mutation analysis. We have introduced several modifications to the SSCP method, which overcome the problem of incomplete denaturation or reannealing of DNA during electrophoresis. The modifications consist of asymmetrical PCR amplification of the sequence of interest, electrophoresis with a higher concentration of acrylamide, and the analysis of the DNA fragments under u.v. light. We have applied this method to the analysis of two specific diseases: neurofibromatosis type 1 (NF1) and cystic fibrosis (CF) from PCR amplified exons. Two single nucleotide changes were observed with this method.",
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AB - The Single Strand Conformation Polymorphism (SSCP) technique is widely used in mutation analysis. We have introduced several modifications to the SSCP method, which overcome the problem of incomplete denaturation or reannealing of DNA during electrophoresis. The modifications consist of asymmetrical PCR amplification of the sequence of interest, electrophoresis with a higher concentration of acrylamide, and the analysis of the DNA fragments under u.v. light. We have applied this method to the analysis of two specific diseases: neurofibromatosis type 1 (NF1) and cystic fibrosis (CF) from PCR amplified exons. Two single nucleotide changes were observed with this method.

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