Ligation-Independent Mechanism of Multiplex Ligation-Dependent Probe Amplification

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  • UNO Naoki
    Department of Laboratory Medicine, Nagasaki University Graduate School of Biomedical Sciences
  • YANAGIHARA Katsunori
    Department of Laboratory Medicine, Nagasaki University Graduate School of Biomedical Sciences

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Description

Multiplex ligation-dependent probe amplification (MLPA) is a widely used technique for detecting genomic structural variants. The technique is based on hybridization and ligation, followed by amplification of the ligation products. Therefore, ligation is considered a fundamental process that determines the feasibility and fidelity of MLPA. However, despite the widespread use of this technique, its reaction mechanism has not been fully analyzed. Herein, we describe a ligation-independent pathway for MLPA and introduce a ligation-independent probe amplification system that can be used to obtain amplified products without the hybridization and ligation processes. Fragment analysis revealed that the ligation-independent pathway is functional and that the capacity to discriminate single nucleotides with MLPA does not depend on ligation. These findings indicate that the feasibility and fidelity of MLPA do not rely on ligation.

Journal

  • Analytical Sciences

    Analytical Sciences 30 (8), 805-810, 2014

    The Japan Society for Analytical Chemistry

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