Analytical Sciences
Online ISSN : 1348-2246
Print ISSN : 0910-6340
ISSN-L : 0910-6340
Original Papers
Ligation-Independent Mechanism of Multiplex Ligation-Dependent Probe Amplification
Naoki UNOKatsunori YANAGIHARA
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Supplementary material

2014 Volume 30 Issue 8 Pages 805-810

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Abstract

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.

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© 2014 by The Japan Society for Analytical Chemistry
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