Multiplex Ligation-Dependent Probe Amplification (MLPA)
Multiplex Ligation-dependent Probe Amplification (MLPA) is a multiplex PCR method detecting abnormal copy numbers of specific genes, including small intragenic rearrangements for up to 50 different genomic DNA or RNA sequences. MLPA is a very important technique in the assessment of hereditary diseases including cancers. It is especially indicated whenever large deletions or duplications are described for a gene.
What are the advantages of MLPA?
• Methods which were primarily developed for detecting point mutations (such as PCR sequencing) generally fail to detect copy numbers changes
• Southern blot analysis will not always detect small deletions and is not ideal as a routine technique
• Although well-characterised deletions and amplifications can be detected by PCR, the exact breakpoint site of most deletions is unknown
• As compared to Fluorescence In Situ Hybridization (FISH), MLPA not only has the advantage of being a multiplex technique, but also one in which very small (50-70 nt) sequences are targeted, enabling MLPA to identify the frequent, single gene aberrations which are too small to be detected by FISH
• MLPA is relatively a low cost and technically uncomplicated method
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