A fast polymerase chain reaction-mediated strategy for introducing repeat expansions into CAG-repeat containing genes.
We describe a simple method for expanding CAG trinucleotides in CAG-repeat containing genes which, in contrast to other techniques, leaves the original gene sequence intact. Here, we expanded the CAG stretches of a plasmid clone containing the cDNA of the SCA3/MJD gene from 22 CAG up to > 130 CAG repeats using polymerase chain reaction (PCR)-mediated mutagenesis. The method reported in this article requires minimal resources, is very fast, and enables to construct recombinant plasmids carrying large CAG expansions.
Laccone
F
F
Institute of Human Genetics, Faculty of Medicine, University of Göttingen, Germany. flaccon@gwdg.de
Maiwald
R
R
Bingemann
S
S
D001483:Base Sequence; D006801:Humans; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*; D019680:Trinucleotide Repeat Expansion