Capillary zone electrophoresis in polymer networks of polymerase chain reaction-amplified oligonucleotides: the case of congenital adrenal hyperplasia.
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 microM 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 comparable 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.
Faculty of Pharmacy, University of Milan, Italy.
C
C
Gelfi
A
A
Orsi
P G
PG
Righetti
M
M
Zanussi
P
P
Carrera
M
M
Ferrari
D000312:Adrenal Hyperplasia, Congenital / Q000235:genetics*; D001483:Base Sequence; D002197:Capillary Action; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004996:Ethidium; D017353:Gene Deletion; D006801:Humans; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000032:analysis* / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011108:Polymers; D013255:Steroid 21-Hydroxylase / Q000737:chemistry / Q000235:genetics*
D009841:Oligonucleotides; D011108:Polymers; D004247:DNA; D013255:Steroid 21-Hydroxylase; D004996:Ethidium