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SALSA MLPA KIT P017 MEN1
Please note that the P244 MEN1-AIP probemix contains the same MEN1 probes as this P017 probemix, supplemented with probes for the AIP gene and several other genes in the 11q13 region. Multiple endocrine neoplasia is an autosomal dominant disorder characterized by a high frequency of peptic ulcer disease and primary endocrine abnormalities involving the pituitary, parathyroid, and pancreas. Type 1 MEN is the result of defects in the Menin protein that is coded by the MEN1 gene. MEN1 is a relatively small gene containing 10 exons in a 7 Kb chromosomal region on chromosome 11q13. The loss of heterozygosity (LOH) that is frequently observed in MEN1 tumors suggests that MEN1 acts as a tumor suppressor gene, as postulated by the Knudson 2-hit hypothesis. In addition to point mutations, several deletions involving one or more complete exons in the MEN1 gene have been described. A deletion of one or more exons of a gene will usually not be detected by techniques like sequencing or DGGE, as a normal copy is also present. FISH is useful in case the complete gene area is deleted but is not sensitive enough to detect copy number changes of a single exon. Southern blots will not detect deletions that extend beyond the probe sequence and are difficult to perform quantitatively. Real time PCR is not reproducible enough to reliably detect a 50% decrease in copy number of a small genomic sequence, and cannot be easily used in multiplex format. MLPA is a useful alternative to detect copy number changes of part, or the complete MEN1 gene. The P017 probe mix included in this MLPA kit contains probes for 7 of the 10 exons of the MEN1 gene on 11q13, two probes for sequences at a rather close distance to MEN1 as well as 15 control probes for sequences located in other genes. We did not succeed in preparing probes for exons 4, 5 and 6. Distances between the MEN1 exons are however very small. This SALSA MLPA kit is designed to detect deletions/duplications of one or more exons of MEN1. 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 of a single exon therefore always require confirmation by other methods. We have no information on what percentage of defects in these genes is caused by deletions/duplications of complete exons. Please note that most defects in these genes are expected to be small (point) mutations, most of which will not be detected by this MLPA test.
Full mix description (pdf)
Last change in probe mix content: Lot 0507 (July 2007) Current Lot Number.: Lot 0507
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
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