Capillary zone electrophoresis in polymer networks of polymerase chain reaction-amplified oligonucleotides: the case of congenital adrenal hyperplasia

Cecilia Gelfi, Antonia Orsi, Pier Giorgio Righetti, Monica Zanussi, Paola Carrera, Maurizio Ferrari

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Abstract

The use of capillary zone electrophoresis (CZE) in polymer networks for the analysis of an 8 bp (base pair) deletion in congenital adrenal hyperplasia was investigated. Separations were performed in Tris-borate-EDTA buffer (pH 8.3) containing 6% liquid linear polyacrylamide as a sieving dynamic matrix and 10 μM ethidium bromide for improving DNA fragment separation. Easy analysis and detection of the 127 and 135 bp amplified fragments was accomplished. The capillary column can be used for >50 analyses before degradation and loss of resolution. The results are compatible to those obtained by gel-slab zone electrophoresis in a 12%T, 4%C polyacrylamide matrix. The sensitivity, by simple UV absorption at 254 nm, is similar to that obtained in gel slabs by dye intercalation staining.

Original languageEnglish
Pages (from-to)201-205
Number of pages5
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume657
Issue number1
DOIs
Publication statusPublished - Jul 1 1994

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Polymerase chain reaction
Electrophoresis
Oligonucleotides
Polymers
Gels
Ethidium
Intercalation
Coloring Agents
Degradation
DNA
Liquids
polyacrylamide
Tris-borate-EDTA buffer

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

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abstract = "The use of capillary zone electrophoresis (CZE) in polymer networks for the analysis of an 8 bp (base pair) deletion in congenital adrenal hyperplasia was investigated. Separations were performed in Tris-borate-EDTA buffer (pH 8.3) containing 6{\%} liquid linear polyacrylamide as a sieving dynamic matrix and 10 μM ethidium bromide for improving DNA fragment separation. Easy analysis and detection of the 127 and 135 bp amplified fragments was accomplished. The capillary column can be used for >50 analyses before degradation and loss of resolution. The results are compatible to those obtained by gel-slab zone electrophoresis in a 12{\%}T, 4{\%}C polyacrylamide matrix. The sensitivity, by simple UV absorption at 254 nm, is similar to that obtained in gel slabs by dye intercalation staining.",
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T2 - the case of congenital adrenal hyperplasia

AU - Gelfi, Cecilia

AU - Orsi, Antonia

AU - Righetti, Pier Giorgio

AU - Zanussi, Monica

AU - Carrera, Paola

AU - Ferrari, Maurizio

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