We recommend confirming all abnormalities detected by MLPA or digitalMLPA whenever possible. You can either use a different (confirmation) probemix when available or an independent technique.

Single-probe aberrations

It is essential to confirm copy number changes detected by a single probe. Single probes may more readily be affected by experimental issues or variability. In addition, a mutation or polymorphism in the probe's target sequence may lead to a reduced signal, which can even mimic a deletion. Sanger sequencing of the probe's target sequence can reveal if there are any variants in the probe's target sequence that could explain a reduced signal.

Confirmation probemixes

If available, you can use a specifically designed confirmation probemix to confirm results from initial testing. A confirmation probemix targets locations close to the targets of the probemix intended for initial testing, but with sufficient distance between ligation sites to allow confirmation of results. You should not use confirmation probemixes for initial testing.

The existence of a confirmation probemix is clearly indicated on product pages and in product descriptions.

Different techniques

It may be possible to confirm results obtained with digitalMLPA with an MLPA probemix, or vice versa. If this is possible, this is usually mentioned in the product documentation.

You can also use a variety of other techniques to confirm results, including Sanger sequencing of (long-range) PCR products, qPCR, long-range PCR, array-CGH and various NGS techniques. The method that is most suitable depends on e.g. the complexity of the region, the size of the aberration, and the availability of other techniques.