Development of RNA-SSCP protocols for the identification and screening of CFTR mutations: Identification of two new mutations

L. Bisceglia, A. Grifa, L. Zelante, P. Gasparini

Research output: Contribution to journalArticle

Abstract

A strategy is described that allows a rapid and accurate identification and screening of cystic fibrosis gene mutations. It consists of setting up and developing RNA single strand conformation polymorphism (rSSCP) protocols, a technique based on the large repertoire of secondary structure of single- stranded RNA. By incorporating the T7 phage promoter sequence into PCR primers, it is possible to carry out rSSCP and compare it to standard single- strand conformation polymorphism (SSCP). Several parallel tests indicate that rSSCP detects a higher fraction of single base changes, and is less time consuming than SSCP since it requires only one fairly short electrophoretic run. Using this technique we were able to identify two new splicing mutations in introns 5 (711 + 5G→A) and 10 (1717-8G→A) of the CFTR gene.

Original languageEnglish
Pages (from-to)136-140
Number of pages5
JournalHuman Mutation
Volume4
Issue number2
Publication statusPublished - 1994

Keywords

  • Cystic fibrosis
  • DNA mutations
  • RNA-SSCP
  • SSCP

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

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