Identification ofCYP21 mutations, one novel, by single strand conformational polymorphism (SSCP) analysis

Detection of Mutations by Single-strand Conformational Polymorphism.

MATERIALS 10x Amplification buffer Include 0.01% (w/v) gelatin in the buffer. 10x TBE electrophoresis buffer Use TBE at a working strength of 1x (89 mM Tris-borate, 2 mM EDTA) for polyacrylamide gel electrophoresis. 6x Gel-loading buffer I Formamide loading buffer Human genomic DNA to be screened for point mutations Dissolve the DNA at 10 μg/ml in TE (pH 7.6). Oligonucleotide primers, forward a...

Screening for mutations by RNA single-strand conformation polymorphism (rSSCP): comparison with DNA-SSCP.

Single-strand conformation polymorphism (SSCP) is a simple method for detecting the presence of mutations in a segment of DNA, but the fraction of all mutations detected is unclear. We have evaluated SSCP for the detection of single-base mutations in the factor IX gene. Multiple conditions were examined including electrophoresis temperature, electrophoresis buffer concentration, acrylamide to b...

F-SSCP: fluorescence-based polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis.

A fluorescence-based method for polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis, F-SSCP, was developed in which the target sequence is amplified by the PCR using fluorescent primers. The amplified products are then heat-denatured and applied to a water-jacket controlled gel in an automated DNA sequencer. The separated strands are detected as laser-excited f...

Identification of Polymorphism in the Keratin Genes (KAP3.2, KAP6.1, KAP7, KAP8) and Microsatellite BfMS in Merino Sheep Using Polymerase Chain Reaction-Single Strand Conformational Polymorphism (PCR-SSCP) Analysis

Determination of apolipoprotein E genotypes by single-strand conformational polymorphism.

We used single-strand conformational polymorphism (SSCP) to determine apolipoprotein E (Apo E) genotypes in 47 individuals. A 295-base-pair (bp) DNA fragment coding for amino acid residues 80-178 of the Apo E protein gave distinct patterns for the three alleles. When we used SSCP to determine the Apo E polymorphism of five individuals whose phenotyping results differed from those of genotyping,...