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Triblock copolymer‐based microchip device for rapid analysis of stuffer‐free multiplex ligation‐dependent probe amplification products
Author(s) -
Shin Gi Won,
Kim Yong Tae,
Heo Hyun Young,
Chung Boram,
Seo Tae Seok,
Jung Gyoo Yeol
Publication year - 2012
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201200436
Subject(s) - multiplex ligation dependent probe amplification , multiplex , computer science , biology , bioinformatics , genetics , gene , exon
Recent improvements in the multiplex ligation‐dependent probe amplification ( MLPA ) method promise successful multiplex analysis of various genetic markers. In particular, it has been demonstrated that elimination of the stuffer sequence included in MLPA probes for length‐dependent analysis substantially simplifies the probe design process and improves the accuracy of the analysis. As is the case for other CE ‐based methods, MLPA could be further developed on a microchip platform. However, high‐resolution analysis of short MLPA probes requires careful microchip operation. In this study, we developed a microchip device for the multiplex analysis of five food‐borne pathogens using a stuffer‐free probe set. Microchip channel design and electrophoresis operating conditions were first optimized for reproducible analysis, after which two sieving matrices were tested. Finally, the method was validated using DNA samples isolated from intentionally infected milk.