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There are a number of techniques available for the detection and characterization of mutations that take advantage of the flexibility and power of polymerase chain reaction (PCR) (Table 1 ). Sequencing remains the gold standard, but it is not very efficient as a screening tool, in particular when evaluating large numbers of samples, or multiple or extended stretches of DNA. f the available techniques, single-strand conformation polymorphism analysis (SSCP) appears as one of the most versatile techniques, amenable to automation, and optimal for screening for known or unknown mutations.
Table 1Main PCR-Based Strategies for Detection of Mutations
Technique | Relevant reference |
|---|---|
Sequencing of PCR products | (16) |
Mutation-specific priming | (17) |
Restriction-enzyme analysts of PCR products | (18) |
Selective ohgonucleotide hybridization | (19) |
Constant denaturmg gel electrophoresis (CDGE) | (20) |
Temperature gradient gel electrophoresis (TGGE) | (21) |
Dtdeoxy tingerprmting | (22) |
Heteroduplex formation analysis | (23) |
Cleavase fragment-length polymorphism (CFLP) | (24) |
Ribonuclease A cleavage of mismatched RNA. DNA duplexes | (25) |
Single-strand conformation polymorphism (SSCP) | (1) |
