Event
Bioengineering Seminar Series: Gyoo Yeol Jung
Friday, September 3, 2010
11:00 a.m.-12:00 p.m.
Room 2108, Chemical and Nuclear Engineering Bldg.
Professor Joonil Seog
jseog@umd.edu
High Fidelity Genetic Analysis System Based on Single-Strand Conformation Dependent Capillary Electrophoresis
Gyoo Yeol Jung
Department of Chemical Engineering
School of Interdisciplinary Bioscience & Bioengineering
Pohang University of Science and Technology, Korea
Molecular diagnostics is a new area of clinical diagnosis, and it includes all tests and methods to identify a disease or the predisposition for a disease analyzing DNA, RNA or proteins of an organism. After completion of the human genome project, enormous reports on genetic variations were reported which cause phenotypic variations and disease conditions. DNA microarray, which can detect thousands of DNA sequences simultaneously, was the major technology to discover a novel DNA sequence area which could differentiate between one condition and another; the DNA sequence is generally called genetic marker. After the genetic marker discovery, validation by quantitative method such as real-time qPCR is often a required step because of error-prone nature of DNA microarray. For molecular diagnosis using discovered genetic markers, however, both DNA microarray and real-time qPCR are not appropriate because DNA microarray is error-prone by nature and real-time qPCR cannot detect multiple targets by limitation on fluorescence dye.
Capillary electrophoresis-based single-strand DNA conformation polymorphism analysis (CE-SSCP), which separates DNA molecules by conformational difference of ssDNA and quantifies them by fluorescence intensity, can detect multiple genetic markers quantitatively. Recently, multiple ligation dependent probe amplification (MLPA) has developed to amplify multiple genetic markers linearly and minimize false positive signal due to the additional ligation step of two probes of a single target. MLPA is, however, confined with the addition of stuffer sequence for the separation step after amplification and only a limited application is available thus far. In this study, we developed a sensitive CE-SSCP system using a novel polymer gel matrix and consequently stuffer-free MLPA system could be developed based on this high-resolution CE-SSCP system. Examples on CNV, pathogen quantification, and transcript profiling will illustrate the potentials of this system as a molecular diagnosis system.
