NONISOTOPIC DETECTION OF MUTATIONS USING A MODIFIED SINGLE-STRAND CONFORMATION POLYMORPHISM ANALYSIS

This report describes a rapid convenient screening system with improve d sensitivity to detect mutations in the cystic fibrosis transmembrane regulator (CFTR) gene based on nonisotopic SSCP analysis, Because con ventional SSCP analysis is often hampered by poor yield of single-stra nded DNA, we applied the well established solid phase technique in whi ch streptavidin-coated magnetic beads are used to immobilize biotinyla ted polymerase chain reaction (PCR) products. High yield of single-str anded DNA can be eluted from the solid phase by denaturation and used for SSCP analysis, An additional advantage of this procedure is that t he immobilized single strand is available without any further purifica tion steps for solid-phase sequencing. (C) 1994 Wiley-Liss, Inc.