CAG triplet analysis in families with androgen insensitivity syndrome by capillary electrophoresis in polymer networks

Cecilia Gelfi, Pier Giorgio Righetti, Fiorella Leoncini, Valeria Brunelli, Paola Carrera, Maurizio Ferrari

Research output: Contribution to journalArticle

Abstract

The potential use of capillary zone electrophoresis in polymer networks (linear polymers above the entanglement threshold, added to the background electrolyte for sieving purposes) for analysis of DNA fragments amplified by a polymerase chain reaction, is shown. In typical runs, the capillary is filled with Tris-borate-EDTA buffer, at pH 8.3, containing 6% linear polyacrylamide as a dynamic sieving matrix. Such formulations allow replenishing the capillary with fresh sieving solution when resolution decays after prolonged use (typically >30 injections per capillary are obtained). The DNA fragments are detected by their intrinsic absorbance at 254 nm. This system has been applied to the analysis of CAG triplet polymorphism in families carrying the androgen insensitivity syndrome. While easy separation is obtained for fragments 139 base pairs (bp) and 160 bp (in families carrying a difference of 7 CAG repeats) even more difficult cases (such as those of families exhibiting fragments of 136 and 139 bp, thus differing by only one CAG repeat) are resolved with precision and diagnostic value.

Original languageEnglish
Pages (from-to)463-468
Number of pages6
JournalJournal of Chromatography A
Volume706
Issue number1-2
DOIs
Publication statusPublished - Jul 7 1995

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ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry
  • Organic Chemistry

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