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SALSA MS-MLPA KIT ME011 Mismatch repair genes
[http://www.geneclinics.org/profiles/hht/details.html]
The Mismatch Repair (MMR) system is critical for the maintenance of genomic stability. MMR increases the fidelity of DNA replication by identifying and excising single-base mismatches and insertion-deletion loops that may arise during DNA replication. Cells with MMR deficiency may lead to the accumulation of mutations resulting in the initiation of cancer. The MMR genes are involved in one of the most prevalent cancer syndromes in humans known as hereditary nonpolyposis colon cancer (HNPCC). There are several proteins required for the complete MMR system. Mutations in the MLH1 and MSH2 have been found in about 90% of HNPCC cases. Mutations in other MMR genes have been less frequent in HNPCC patients. In many sporadic colon cancers, hypermethylation of the MLH1 gene promoter resulting in its transcriptional silencing has been observed more than mutations. This ME011 MMR probemix has been developed to detect aberrant CpG islands methylation of six MMR genes and includes 5 probes for MLH1, 3 probes MSH2, 3 probes for MHSH6, 3 probes for MSH3, 1 probe for MLH3, 3 probes for PMS2 and 3 probes specific for the MGMT promoter region. MGMT plays a role in removing O(6)-alkylguanine which is the major mutagenic and carcinogenic lesion induced by alkylating mutagens. The MLH1 gene is located at chromosome 3p22.1, MLH3 at chromosome 14q24.3, MSH2 at 2p21, MSH3 at 05q14.1, MSH6 gene at 2p16, PMS2 at 7p22 and the MGMT gene is located at chromosome 10q26. This kit includes 21 probes that contain a HhaI recognition site, which yields information about the methylation status of a target sequences. In addition, 11 different control probes are present which are not influenced by the HhaI digestion. Besides the detection of aberrant methylation, all probes present will give information on copy number changes in the analysed sample. This MS-MLPA kit is designed to detect and quantify abnormal methylation ratios of MSH2, MLH1, MLH3, PMS2, MSH3, MSH6, MGMT, as well as deletions and duplications of one or more exons of the genes. Deletions of probe recognition sequences will be apparent by a 35-50% reduced relative peak area of the amplification product of that probe. However, mutations/polymorphisms very close to the probe ligation site may also result in a reduced relative peak area. Apparent deletions detected by single probe will therefore always require confirmation by other methods.
Full mix description (pdf)
Last change in probe mix content: Lot 0407 (April 2007) Current Lot Number.: Lot 0408
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 - MS-MLPA: an attractive alternative laboratory assay for robust, reliable, and semiquantitative detection of MGMT promoter hypermethylation in gliomas.
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