title: Precise expression profiling by stuffer-free multiplex ligation-dependent probe amplification.
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.
authors.affiliation: Institute of Environmental and Energy Technology, Pohang University of Science and Technology , Pohang, Gyeongbuk 790-784, Korea.
authors.forename: Gi Won
authors.initials: GW
authors.lastname: Shin
authors.forename: Jeongkyeong
authors.initials: J
authors.lastname: Na
authors.forename: Mihwa
authors.initials: M
authors.lastname: Seo
authors.forename: Boram
authors.initials: B
authors.lastname: Chung
authors.forename: Hong Gil
authors.initials: HG
authors.lastname: Nam
authors.forename: Seung-Jae
authors.initials: SJ
authors.lastname: Lee
authors.forename: Gyoo Yeol
authors.initials: GY
authors.lastname: Jung
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics* / Q000378:metabolism; D019075:Electrophoresis, Capillary; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D060885:Multiplex Polymerase Chain Reaction