Detection of point mutations in the p53 gene: comparison of single-strand conformation polymorphism, constant denaturant gel electrophoresis, and hydroxylamine and osmium tetroxide techniques.
A comparison was made between the 3 most commonly used techniques for the detection of point mutations: single-strand conformation polymorphism (SSCP), constant denaturant gel electrophoresis (CDGE), and hydroxylamine and osmium tetroxide used in amplification mismatch cleavage analysis (HOT). Using human DNA samples containing known mutations in the p53 gene, SSCP detected 90% of mutations (18/20), CDGE detected 88% (15/17) pre-decoding of the samples but 100% when the mutations were known and the CDGE conditions optimized, and the HOT technique was 100% accurate, although 1 mutation was missed through careless examination of the gel. The positive and negative aspects of each of the techniques are considered and suggestions are made regarding the particular situations in which each of them is most useful.
Condie
A
A
Human Genetics Unit, Western General Hospital, Edinburgh, Scotland.
Eeles
R
R
Borresen
A L
AL
Coles
C
C
Cooper
C
C
Prosser
J
J
D004277:DNA, Single-Stranded / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D016158:Genes, p53; D005821:Genetic Techniques; D019811:Hydroxylamine; D006898:Hydroxylamines; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D009993:Osmium Tetroxide; D017354:Point Mutation; D011110:Polymorphism, Genetic
D004277:DNA, Single-Stranded; D006898:Hydroxylamines; D019811:Hydroxylamine; D009993:Osmium Tetroxide