General information
The SALSA MLPA
Probemix P432 MYH9 is a
research use only (RUO) assay for the detection of deletions or duplications in the
MYH9 gene, which is associated with
MYH9-related disorder (MYH9RD). This probemix can also be used to detect the presence of the c.287C>T (p.S96L) and c.2104C>T (p.R702C) point mutations as well as the wild type sequence of the c.4270G>C/A/T (p.D1424H/N/Y) mutation.
The
MYH9 gene is located on chromosome 22q12.3 and encodes the non-muscle myosin (heavy chain, type II-A).
MYH9-related disorder (MYH9RD) is an autosomal dominant platelet disorder characterized by macrothrombocytopenia and thrombocytopenia. The complex phenotype of MYH9RD may include bleeding problems, hearing loss, renal disease, and cataracts. Epstein syndrome, Fechtner syndrome, May-Hegglin anomaly and Sebastian syndrome, previously described as distinct disorders, represent some of the different clinical presentations of MYH9RD.
More information is available at
https://www.ncbi.nlm.nih.gov/books/NBK2689/.
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 P432-A2 MYH9 contains 53 MLPA probes with amplification products between 128 and 500 nucleotides (nt). This includes 42 probes for the
MYH9 gene. Furthermore, this probemix also contains two probes specific for the c.287C>T and the c.2104C>T mutation, which will only generate a signal when the respective mutation is present. One probe that will only detect the wild type sequence of the c.4270G>C/A/T mutation is also included. In addition, eight reference probes are included 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).
This probemix contains nine quality control fragments generating amplification products between 64 and 105 nt: four DNA Quantity fragments (Q-fragments), two DNA Denaturation fragments (D-fragments), one Benchmark fragment, and one chromosome X and one chromosome Y-specific fragment. More information on how to interpret observations on these control fragments can be found in the MLPA General Protocol and online at
www.mrcholland.com.
SALSA Binning DNA SD035
The SD035 Binning DNA provided with this probemix can be used for binning of all probes including the two mutation-specific probes (MYH9 probe 18610-SP0671-L24707 for the c.2104C>T mutation, and MYH9 probe 18616-SP0675-L24711 for the c.287C>T mutation). SD035 Binning DNA is a mixture of genomic DNA from healthy individuals and synthetic DNA that contains the target sequence detected by the above mentioned probe. Inclusion of one reaction with 5 μl SD035 Binning DNA in initial MLPA experiments is essential as it can be used to aid in data binning of the peak pattern using Coffalyser.Net software. Furthermore, Binning DNA should be included in the experiment whenever changes have been applied to the set-up of the capillary electrophoresis device (e.g. when capillaries have been renewed). Binning DNA should never be used as a reference sample in the MLPA data analysis, neither should it be used in quantification of mutation signal(s). It is strongly advised that all samples tested are extracted with the same method and derived from the same source of tissue. For further details, please consult the SD035 Binning DNA product description, available online:
www.mrcholland.com.
This product is for research use only (RUO).
Sample DNA
Sample DNA developed for this product:
The product description of this Sample DNA can be found on this page under
Online available downloads >
Other product documentation.