General information
The SALSA MLPA
Probemix P300 Reference-2 is a
research use only (RUO) assay which is developed to be used in combination with ‘home-made’ synthetic (MS-)MLPA probes. This probemix contains reference probes and control fragments specific for unique human DNA sequences. The control fragments and reference probes in this P300 probemix facilitate the data analysis of a synthetic probemix with home-made probes.
MRC Holland also offers the SALSA MLPA
Probemix P200 Reference-1. While the P200 Reference-1 probemix leaves the entire range from 80-170 nt open for the inclusion of synthetic probes, this P300 Reference-2 probemix has reference probes distributed over the whole size range of the probemix. This is especially advantageous for correction of the signal sloping of (MS-)MLPA amplification products. Signal sloping is the effect that longer probes generate a lower peak height/area on the electropherogram than shorter probes. The exact amount of signal sloping differs between sequencer types and can differ between different samples.
The use of P200 is recommended when a large number of synthetic probes are used, preferably targeting sequences on different chromosomes. Results can be difficult to interpret when P200 is used in combination with a small number of probes that all target the same gene or chromosomal region, or with a small number of probes with lengths much shorter than the reference probes. The use of P300 is recommended when a smaller number of synthetic probes is used. Please note that MRC Holland cannot offer support for custom probe design anymore.
This SALSA MLPA probemix is not CE/FDA registered for use in diagnostic procedures. Purchase of this product includes a limited license for research purposes.
Probemix content
The SALSA MLPA Probemix P300-B1 Reference-2 contains 25 MLPA probes with amplification products between 64 and 274 nt. This includes 14 reference probes that detect autosomal chromosomal locations. Complete probe sequences and the identity of the genes detected by the reference probes are available online (
www.mrcholland.com). In addition, four DNA Quantity Fragments (Q-fragments), two DNA Denaturation Fragments (D-fragments), two HhaI-digestion control fragments, one chromosome X and one chromosome Y-specific fragment are included. The Q-fragments give a warning when insufficient amounts of sample DNA are used. The D-fragments give a warning when sample DNA was not completely denatured. More information on how to interpret observations on these control fragments can be found in the (MS-)MLPA General Protocol and online at
www.mrcholland.com.