Title of article :
Direct fluorescence detection of point mutations in human genomic DNA using microbead-based ligase chain reaction
Author/Authors :
Meng، نويسنده , , Xiangxian and Yang، نويسنده , , Xiaohai and Wang، نويسنده , , Kemin and Guo، نويسنده , , Qiuping and Tan، نويسنده , , Yongjun and Mo، نويسنده , , Qiuhua and Xu، نويسنده , , Xiangmin، نويسنده ,
Issue Information :
ماهنامه با شماره پیاپی سال 2010
Pages :
5
From page :
1725
To page :
1729
Abstract :
This report has described a convenient genotyping method capable of detecting point mutations directly in human genomic DNA based on the combination of ligase chain reaction (LCR) and microbead-enrichment technique. LCR primers, including a biotin-labeled common primer and two fluorescence-labeled allele-specific primers, are designed for two alleles of a mutated site. When genomic DNA carries the mutated site, the common primer and allele-specific primer are ligated to form exponential amplified biotin-labeled fluorescence ligation products. These ligated products are enriched by streptavidin-coated microbeads, and genotypes are identified conveniently according to the fluorescence color of microbeads using fluorescent microscopy. Due to amplification of LCR process and enrichment of microbeads, the detection limit of the proposed method is as low as 10−15 mol/L templates. The method provides a convenient and simple strategy to detect point mutation directly in human genome. We have confirmed the efficiency of this approach with the identification of β-globin gene point mutation, which results in the reduced production of globin in an inherited hemoglobin disorder thalassemia disease.
Keywords :
Genotyping , point mutation , Ligase chain reaction , Microbead-enrichment , thalassemia
Journal title :
Talanta
Serial Year :
2010
Journal title :
Talanta
Record number :
1659547
Link To Document :
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