This product is for basic research and intended for experienced users only! Interpretation of results may be complicated, and MRC Holland may only be able to provide basic support.
Discounts on the ME001, ME002, ME003, ME004, ME046 kits are available for specific research projects. Please enquire.
This SALSA® MLPA® probemix is for basic research! This probemix enables you to detect aberrant methylation of CpG islands upstream of genes for which an altered methylation status in one or more types of tumours has been reported in literature. In case interesting results are obtained by users, it is possible to develop methylation probemixes specific for a certain tumour in collaboration with MRC-Holland. Interpretation of results obtained with this product can be complicated. MRC-Holland cannot provide assistance with data interpretation.
Aberrant methylation of CpG-islands has been shown to be associated with transcriptional inactivation of tumour suppressor genes in a wide spectrum of human cancers. CpG islands are located in or near the promoter region or other regulatory regions of approximately 50% of human genes.
This ME001-C2 MS-MLPA probemix contains 26 MS-MLPA probes which detect the methylation status of promoter regions of 24 different tumour suppressor genes. These tumour suppressor genes are frequently silenced by methylation in tumours, but are unmethylated in blood-derived DNA of healthy individuals. In addition, 15 reference probes are included which are not affected by HhaI digestion. Besides detecting aberrant methylation, all 41 probes will give information on copy number changes in the analysed sample. The MLPA reaction requires as little as 20 ng of human DNA and can be used on a variety of DNA samples, including those derived from paraffin-embedded tissues.
The MS-MLPA probes in this ME001-C2 probemix detect sequences in promoter regions of tumour suppressor genes that are unmethylated in most blood-derived DNA samples. Upon digestion, the peak signal obtained in unmethylated samples will be very small or absent. In contrast, when tested on in vitro methylated human DNA, these probes do generate a signal. We have no data showing that methylation detected by a particular probe indeed influences the corresponding mRNA level.
This SALSA® MS-MLPA® probemix can be used to detect aberrant methylation of one or more sequences of the tumour suppressor genes. Methylation levels can be different for different tissues. If possible, use identically treated test and reference samples (same tissue type and extraction method). This SALSA® MS-MLPA® probemix can be used to detect deletions/duplications of one or more sequences in the above mentioned chromosomal regions in a DNA sample. Heterozygous deletions of recognition sequences should give a 35-50% reduced relative peak height of the amplification product of that probe. Note that a mutation or polymorphism in the sequence detected by a probe can also cause a reduction in relative peak height, even when not located exactly on the ligation site! In addition, some probe signals are more sensitive to sample purity and small changes in experimental conditions. Therefore, deletions and duplications detected by MLPA should always be confirmed by other methods. Not all deletions and duplications detected by MLPA will be pathogenic; users should always verify the latest scientific literature when interpreting their findings. Finally, note that most defects in this gene are expected to be small (point) mutations which will not be detected by this SALSA® MS-MLPA® test. We have no information on what percentage of defects in these genes is caused by deletions/duplications of complete exons.