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RESEARCH PRODUCT

Multiplex Ligation-dependent Probe Amplification (MLPA)

Rosa NogueraAna P. BerbegallSamuel NavarroEva Villamón

subject

musculoskeletal diseasescongenital hereditary and neonatal diseases and abnormalitieschemistry.chemical_compoundgenomic DNAchemistryHybridization probeCritical factorsPcr cloningMultiplexComputational biologyMultiplex ligation-dependent probe amplificationDNA extractionDNA

description

The Multiplex Ligation-dependent Probe Amplification (MLPA) is a PCR-based method. The procedure relies on sequence-specific probe hybridization of genomic DNA, followed by multiplex-PCR amplification of the hybridized probe and a semiquantitative analysis of the resulting PCR products. MLPA allows the analysis of around 40 loci in the same reaction, and is a sensitive and relatively fast technique. Only a small amount of DNA is required and results are available within 2 days.The critical factors when performing MLPA analyses from formalin-fixed paraffin-embedded (FFPE) tissues are DNA integrity and purity; for this reason, a suitable DNA extraction method must be chosen.The MLPA protocol described in this chapter provides a methodology for analyzing the genetic copy number changes from FFPE tissues using MLPA P-products (MRC-Holland). The steps include DNA extraction from tissue sections, DNA sample denaturation, and MLPA data analysis.

yearjournalcountryeditionlanguage
2011-01-01
https://doi.org/10.1007/978-3-642-17890-0_33