PCR products with heterozygous mutations containing two types of heteroduplexes.
- Author:
Xian-ning ZHANG
1
;
Xin-hui HE
;
Ji-cheng LI
Author Information
- Publication Type:Journal Article
- MeSH: Base Pair Mismatch; Chromatography, High Pressure Liquid; DNA Mutational Analysis; Electrophoresis, Polyacrylamide Gel; Heteroduplex Analysis; Heterozygote; Humans; Keratin-9; Keratins; genetics; Mutation; Nucleic Acid Heteroduplexes; Polymerase Chain Reaction
- From: Journal of Zhejiang University. Medical sciences 2005;34(5):417-420
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo confirm that PCR products with heterozygous mutations contain not only wide-type and mutant homoduplexes, but also two types of heteroduplexes.
METHODSAn insertion-deletion mutation in the exon 1 of KRT9 gene (497delAinsGGCT), which caused Chinese epidermolytic palmoplantar keratoderma (EPPK) was investigated by polymerase chain reaction (PCR), polyacrylamide gel electrophoresis (PAGE) and denaturing high-performance liquid chromatography(DHPLC).
RESULTSTwo heteroduplexes and two homoduplexes in the PCR product from the heterozygous mutation of the exon 1 of KRT9 (497delAinsGGCT) were detected.
CONCLUSIONPCR products from KRT9 gene with heterozygous mutations contain two types of heteroduplexes. It is without the need to perform heating and cooling PCR products obtained from heterozygous mutations in advance before the mutation screening steps such as denaturing gradient gel electrophoresis (DGGE), temperature gradient gel electrophoresis (TGGE), conformation-sensitive gel electrophoresis (CSGE), DHPLC and heteroduplex analysis (HA), etc.