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Precise Expression Profiling by Stuffer-Free Multiplex Ligation-Dependent Probe Amplification

Title
Precise Expression Profiling by Stuffer-Free Multiplex Ligation-Dependent Probe Amplification
Authors
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
2013-10-01
Citation
Analytical Chemistry, 85(19), 9383-9389
Type
Article
Article Type
Article
Keywords
Alternative TechnologiesAnimalAnimalsBiological StudiesCaenorhabditis ElegansCapillary ElectrophoresisElectrophoresis, CapillaryEscherichia ColiExpression ProfilingGain ControlGene ExpressionGeneticsMetabolic GenesMetabolismMultiplex AnalysisMultiplex Polymerase Chain ReactionMultiplexingPolymerase Chain ReactionProbe AmplificationProbesProduct DesignReal-Time Polymerase Chain Reaction
ISSN
0003-2700
Abstract
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.
URI
http://hdl.handle.net/20.500.11750/1694
DOI
10.1021/ac402314h
Publisher
American Chemical Society
Related Researcher
Files:
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Collection:
New BiologyETC1. Journal Articles


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