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SALSA MS-MLPA KIT ME003 Tumorsuppressor 3
[http://www.geneclinics.org/profiles/hht/details.html]
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 ME003-A1 MS-MLPA probemix contains 27 MS-MLPA probes which detect the methylation status of promotor regions of 18 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, 13 reference probes are included which are not affected by HhaI digestion. Besides detecting aberrant methylation, all 40 probes present will give information on copy number changes in the analyzed 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. Please note that each MS-MLPA reaction generates two samples that need analysis by capillary electrophoresis: one undigested sample for copy number detection and one digested sample for methylation detection. More information about MS-MLPA can be found on page 2 and in the MS-MLPA protocol. The MS-MLPA probes in this ME003-A1 probemix detect sequences in promoter regions of tumor 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 levels. Moreover, most MS-MLPA probes target only a single specific HhaI site of the CpG island. If methylation is absent for this particular CpG-site, it does not necessarily mean that the complete CpG island is unmethylated. For samples containing both tumor and normal cells, MLPA will indicate the average copy number of genes and the average methylation level.
Full mix description (pdf)
Last change in probemix content : Lot 0608 (June 2008) Current lot number : Lot 0608
IMPORTANT NOTICE: MLPA kits are sold by MRC-Holland for research purposes and to demonstrate the possibilities of the MLPA technique. This kit is not CE/FDA certified for use in diagnostic procedures. Salsa MLPA kits are supplied with all necessary buffers and enzymes. Purchase of the Salsa MLPA test kits includes a limited license to use these products for research purposes. The use of this MLPA kit requires a thermocycler with heated lid and sequence type electrophoresis equipment. Different fluorescent PCR primers are available. The MLPA technique has been first described in Nucleic Acid Research 30, e57 (2002)
References 2007 -- Sequential gene promoter methylation during HPV-induced cervical carcinogenesis. 2006 -- Epigenetic events of disease progression in head and neck squamous cell carcinoma.
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