Capillary electrophoresis of polymerase chain reaction-amplified products in polymer networks: the case of Kennedy's disease.
In Kennedy disease (an X-linked motoneuronal disorder associated with an increase in the number of (CAG)n triplet repeats in the first exon of the Androgen receptor gene; about twice as many as in normal conditions), polymerase chain reaction-amplified genic products exhibit two DNA fragments in the heterozygous female carriers, one with a range between 468 and 495 bp in the normal polymorphic population and a fragment corresponding to the pathological state that reaches 573 bp. These fragments are easily separated by gel-slab electrophoresis and detected by intercalating dye staining (ethidium bromide). As an alternative procedure, capillary zone electrophoresis in polymer networks, consisting of 8% polyacryloylaminoethoxyethanol at 0% cross-linker, offers a simple procedure for separation and on-line detection via UV absorbance at 254 nm, thus avoiding additional staining steps. The capillary column can be repeatedly used for up to 80-100 times and the electropherogram is stored on a magnetic support. Easy comparison among different runs is obtained by aligning all tracings to an internal standard of a 650 bp fragment added as a marker.
Nesi
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Istituto di Chimica degli Ormoni, CNR, Milano, Italy.
Righetti
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PG
Patrosso
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MC
Ferlini
A
A
Chiari
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M
D000180:Acrylic Resins; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005260:Female; D005815:Genetic Code; D006801:Humans; D008297:Male; D009134:Muscular Atrophy, Spinal / Q000175:diagnosis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011944:Receptors, Androgen / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid
D000180:Acrylic Resins; D011944:Receptors, Androgen; C089575:poly(N-acryloylaminoethoxyethanol)