Methylation analysis of hMLH1 gene promoter by a bisulfite-sensitive single-strand conformation polymorphism–capillary electrophoresis method

2006 ◽  
Vol 20 (8) ◽  
pp. 815-820 ◽  
Author(s):  
Xianzhe Shi ◽  
Jianhua Li ◽  
Chunxia Zhao ◽  
Shen Lv ◽  
Guowang Xu
Keyword(s):  
Capillary Electrophoresis ◽  
Gene Promoter ◽  
Single Strand Conformation Polymorphism ◽  
Single Strand ◽  
Methylation Analysis ◽  
Capillary Electrophoresis Method ◽  
Electrophoresis Method ◽  
Hmlh1 Gene ◽  
Conformation Polymorphism

Multiplex Single Strand Conformation Polymorphism Analysis by Capillary Electrophoresis with On-the-Fly Fluorescence Lifetime Detection

2005 ◽  
Vol 59 (3) ◽  
pp. 335-339 ◽  
Author(s):  
Tara Morcone Snyder ◽  
Linda B. McGown
Keyword(s):  
Capillary Electrophoresis ◽  
Fluorescence Lifetime ◽  
Single Strand Conformation Polymorphism ◽  
Single Strand ◽  
Polymorphism Analysis ◽  
Wild Type ◽  
G20210a Mutation ◽  
Prothrombin Gene ◽  
Fluorescence Lifetime Detection ◽  
Conformation Polymorphism

This paper describes the use of on-the-fly fluorescence lifetime detection (OFLD) for multiplex single strand conformation polymorphism (SSCP) analysis by capillary electrophoresis (CE). The dye labels studied for multiplex SSCP-OFLD-CE analyses included RG, NBD, and BODIPY-FL. The dyes were first investigated for a model system of “Wild Type” and “Mutant” 43-base fragments designed to vary by a single A/T substitution. Two dye pairs, BODIPY-FL/RG and BODIPY-FL/NBD, were then used to detect the G20210A mutation in the human prothrombin gene. Mobility correction was required for the BODIPY-FL/RG system. Three “blind” analyses were performed of three mixtures that combined a control fragment (wild type-BODIPY-FL) with two “unknown” fragments selected among four possibilities (wild type or mutant labeled with NBD or RG). In each multiplex analysis, the “origin” of the unknown fragments was correctly identified on the basis of fluorescence lifetime of the dye label and the presence or absence of the mutation was correctly determined on the basis of conformation-induced differences in migration time.

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