Simple detection of tRNA(Lys) mutation in myoclonus epilepsy associated with ragged-red fibers (MERRF) by polymerase chain reaction with a mismatched primer.
We developed a simple method for the detection of a tRNA(Lys) mutation in myoclonus epilepsy associated with ragged-red fibers (MERRF) by polymerase chain reaction with use of a mismatched primer. Although the tRNA(Lys) mutation does not alter recognition sequences for commercially available restriction enzymes, we have successfully changed two nucleotides flanking the A to G mutation at nucleotide position 8344 in a tRNA(Lys) gene of a mitochondrial genome. As a result, the mutation can be detected as a Nae I restriction fragment length polymorphism. With this method, all eight MERRF patients and an asymptomatic mother of a MERRF patient, from six independent families, had the same tRNA(Lys) mutation. Our method is simple and should also be useful for the quantitation of heteroplasmies.
Yoneda
M
M
Department of Neurology, Niigata University, Japan.
Tanno
Y
Y
Nonaka
I
I
Miyatake
T
T
Tsuji
S
S
D001483:Base Sequence; D004272:DNA, Mitochondrial / Q000737:chemistry*; D004831:Epilepsies, Myoclonic / Q000235:genetics*; D005260:Female; D006801:Humans; D008297:Male; D008931:Mitochondria, Muscle / Q000737:chemistry; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012357:RNA, Transfer, Lys / Q000235:genetics*
D004272:DNA, Mitochondrial; D012357:RNA, Transfer, Lys