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SALSA MS-MLPA KIT ME002 Tumorsuppressor 2
[http://www.geneclinics.org/profiles/hht/details.html]
The purpose of this test is to detect aberrant methylation of DNA derived from a variety of samples, including DNA derived from paraffin-embedded tissues. Next to changes in methylation, this test enables detecting copy number changes of 41 loci with the use of only 20 ng of human DNA. Aberrant methylation of CpG-islands has been shown to be associated with transcriptional inactivation of tumor suppressor genes in a wide spectrum of human cancers. CpG-islands are located in or near the promoter region and other regulatory sequences of approximately 50 % of the human genes. This MS-MLPA kit ME002 can be used to detect copy number changes, as well as to analyze CpG islands methylation in a semi-quantitative manner. MLPA probes for methylation quantification are similar to normal MLPA probes, except that the sequence detected by the MS-MLPA probe contains a recognition sequence for the methylation-sensitive restriction enzyme HhaI. Upon digestion of the sample DNA/probe hybrids by HhaI, only probes of which the recognition sequence has been methylated will generate a signal. In contrast, for probes whose HhaI-recognition site is unmethylated, no probe signal will be detected. More information about MS-MLPA can be found on the next pages and in the MS-MLPA protocol. The ME002 MS-MLPA probemix contains 27 sequences that correspond to 25 tumor suppressor genes which are frequently silenced by methylation in different tumors, but are unmethylated in blood-derived DNA of healthy individuals. In addition, 14 different control probes are present that are not influenced by the HhaI digestion. In addition to the detection of aberrant methylation, all 41 probes present will yield information on copy number changes in the analysed sample. This mix can be used as a confirmation of mix ME001B, since ME002 contains almost the same genes as in the ME001B kit (probes for thirteen genes are present in both mixes). The probes in the ME002 kit recognize a different CpG-site as compared to the probes in the ME001B kit!
Full mix description (pdf)
Last change in probemix content : Lot 0706 (July 2006) Current lot number : Lot 1207
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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