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We are glad to inform you that we released an improved version of SALSA® MLPA® Probemix ME029 FMR1-AFF2. ME029 targets the FMR1 and AFF2 genes which contain trinucleotide repeats that are expanded in Fragile X and Fragile XE syndromes. Denaturation of these extremely GC-rich regions requires a product-specific protocol, which has also been updated in the new C1 version.
Left: ME029-B3 supplied with Denaturation Buffer. Right: ME029-C1 supplied with Low pH MLPA Buffer.
The FMR1 and AFF2 regions targeted in SALSA® MLPA® Probemix ME029-C1 FMR1-AFF2 are very difficult to denature, especially in DNA samples containing long trinucleotide expansions. For this reason, an extra Denaturation Buffer was always supplied with previous versions of the ME029 FMR1-AFF2 probemix. However, it was observed that extremely long trinucleotide repeats would not always fully denature even when using the Denaturation Buffer.
To improve this, the use of 60 mM NaOH solution (freshly prepared by the user) now replaces the use of the ME029 FMR1-AFF2 Denaturation Buffer. More details on the NaOH solution can be found on page 2 of the ME029 FMR1-AFF2 product description. At the start of the ME029 MLPA reaction, sample DNA should be mixed with this 60 mM NaOH solution.
As the addition of NaOH to DNA samples increases pH, potentially reducing ligase activity, MRC Holland will include a low-pH version of the MLPA Buffer (purple cap) with each ME029-C1 order to counterbalance this effect. This Low pH MLPA Buffer should always and exclusively be used with the ME029-C1 probemix, instead of the normal MLPA Buffer (yellow cap) that is included in MLPA reagent kits.
We understand that transitioning to a new product version and protocol may require some preparation on your side. Hence, it will be possible to acquire the older ME029-B3 version including the Denaturation Buffer, in reasonable quantities while stocks last, until 29 September 2023. For inquiries regarding ordering the older ME029-B3 version, please contact firstname.lastname@example.org. By default, the new version of the ME029 probemix will be provided if not otherwise specified.