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SALSA MS-MLPA KIT ME001B Tumor suppressor-1
[http://www.geneclinics.org/profiles/hht/details.html]
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 ME001B can be used to detect copy number changes, as well as to analyze CpG islands methylation in a semi-quantitative manner. The purpose of this test is to detect aberrant methylation patterns of DNA derived from a variety of samples including DNA derived from paraffin-embedded tissues. Besides changes in the methylation status, this protocol allows the detection of copy number changes of 40 loci with the use of only 20 ng of human DNA. MLPA probes for methylation quantification are similar to ordinary 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 with HhaI, probes of which the recognition sequence is unmethylated in the sample analysed, will not generate a signal. In contrast, a normal probe signal will be detected if the site is methylated. The ME001B MS-MLPA probemix contains 25 sequences that correspond to a set of tumor suppressor genes that are frequently silenced by methylation in different tumors, but are unmethylated in blood-derived DNA of healthy individuals. In addition 15 different control probes are present that are not influenced by the HhaI digestion. Besides the detection of aberrant methylation, all 40 probes present will yield information on copy number changes in the sample analysed. More information about MS-MLPA can be found on the next page. SALSA MLPA kit ME002 Tumor suppressor-2 can be used to confirm the results of 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 (word)
Full mix description (pdf)
Last change in probe mix content: lot 0407 (April 2007) Current Lot Number.: lot 0407
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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