An assay of gene copy number and its application based on heteroduplex products.
Exp Mol Pathol
; 91(1): 429-33, 2011 Aug.
Article
em En
| MEDLINE
| ID: mdl-21565183
ABSTRACT
The aim of this study was to set up a simple and efficient method for detecting gene copy number, based on heteroduplex products from single-tube PCR/DHPLC. Single-nucleotide polymorphisms (SNPs) on the α-globin gene and chromosome 21 were used as examples. And the formula for quantitative calculation of gene copy number was deduced-based on the peak heights of homoduplexes and heteroduplexes on the DHPLC pattern. 27 samples (14 normal DNA and 13 cases of trisomy-21) were assessed with this method, and 160 samples (48 normal DNA and 112 α-thalassemia samples) were assessed with this method combined with a duplex PCR/DHPLC. Results for 184 of 187 cases were concordant with the known genotypes; three cases of trisomy-21 could not be detected because the target SNPs were homozygous. In conclusion, quantitative assessment of heteroduplex products from single-tube PCR/DHPLC is simple and rapid, and can be used to detect α-thalassemia gene deletions (α(-3.7), α(-4.2)) and trisomy-21.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Bioensaio
/
Cromatografia Líquida de Alta Pressão
/
Dosagem de Genes
/
Análise Heteroduplex
/
Ácidos Nucleicos Heteroduplexes
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
En
Revista:
Exp Mol Pathol
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
China