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Precise Expression Profiling by Stuffer-Free Multiplex Ligation-Dependent Probe Amplification
- Precise Expression Profiling by Stuffer-Free Multiplex Ligation-Dependent Probe Amplification
- Shin, GW[Shin, Gi Won]; Na, J[Na, Jeongkyeong]; Seo, M[Seo, Mihwa]; Chung, B[Chung, Boram]; Nam, HG[Nam, Hong Gil]; Lee, SJ[Lee, Seung-Jae]; Jung, GY[Jung, Gyoo Yeol]
- DGIST Authors
- Nam, HG[Nam, Hong Gil]
- Issue Date
- Analytical Chemistry, 85(19), 9383-9389
- Article Type
- Alternative Technologies; Animal; Animals; Biological Studies; Caenorhabditis Elegans; Capillary Electrophoresis; Electrophoresis, Capillary; Escherichia Coli; Expression Profiling; Gain Control; Gene Expression; Genetics; Metabolic Genes; Metabolism; Multiplex Analysis; Multiplex Polymerase Chain Reaction; Multiplexing; Polymerase Chain Reaction; Probe Amplification; Probes; Product Design; Real-Time Polymerase Chain Reaction
- In systems biological studies, precise expression profiling of functionally important gene sets is crucial. Real-time polymerase chain reaction is generally used for this purpose. Despite its widespread acceptance, however, this method is not suitable for multiplex analysis, resulting in an inefficient assay process. One alternative technology in the spotlight is multiplex ligation-dependent probe amplification (MLPA). But MLPA depends on length-based discrimination of amplified products, which complicates probe design and compromises analysis results. Here, we devised a variation of MLPA that utilizes conformation-sensitive capillary electrophoresis, and demonstrated the simplicity of the probe-design process and improved precision of the assay in analyses of 33 Escherichia coli metabolic genes and 16 Caenorhabditis elegans longevity-related genes. The results showed that relative expression could be quantitatively measured over a relevant dynamic range by using similar-sized probes. Importantly, the improved precision compared to conventional MLPA promises a wider application of this method for various biological systems. © 2013 American Chemical Society.
- American Chemical Society
- Related Researcher
Nam, Hong Gil
CBRG(Complex Biology Research Group)
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