The protein truncation test (PTT) Luijt, Rob B. Van Der; Fodde, Riccardo; Dunnen, Johan T. Den
Mutation Detection,
02/1998
Book Chapter
The detection and characterization of point mutations in genes responsible for inherited disorders has become one of the most common practices in human molecular genetics. Several protocols are ...nowadays available: single strand conformation polymorphism (SSCP) (1), RNase protection (2), hydroxylamine and osmium tetroxide (HOT) chemical cleavage (3), and denaturing gradient gel electrophoresis (DGGE) (4, 5). These techniques are generally aimed at the identification of single base substitutions, insertions, or deletions in relatively small DNA fragments (50-500 bp). However, when dealing with large genes, it is desirable to analyse larger fragments of the coding region to speed up the analysis and pin-point the location of the alteration prior to the nucleotide sequence determination. To this aim, mRNA-based mutation detection protocols are often employed. Moreover, if the spectrum and the type (i.e. missense, nonsense, or frame-shift) of the prevalent mutations at a given diseasecausing gene has already been established, one would prefer to employ a protocol ‘targeted’ for the detection of the most frequent type of mutations.
