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.
Institute of Human Genetics, Faculty of Medicine, University of Göttingen, Germany. flaccon@gwdg.de
F
F
Laccone
R
R
Maiwald
S
S
Bingemann
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