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SALSA MLPA KIT P024-B CDKN2A/2B region
Full mix description (pdf)
Current Lot Number.: Lot 1008
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 2006 -- A novel type of deletion in the CDKN2A gene identified in a melanoma-prone family. Genes Chromosomes Cancer. 2006 -- Fine-mapping loss of gene architecture at the CDKN2B (p15INK4b), CDKN2A (p14ARF, p16INK4a), and MTAP genes in head and neck squamous cell carcinoma. 2005 --Prevalence of 9p21 deletions in UK melanoma families. 2005-- Genome profiling of melanocytic tumors using multiplex ligation-dependent probe amplification (MLPA): Its usefulness as an adjunctive diagnostic tool for melanocytic tumors. 2004-- Measurement of relative copy number of CDKN2A/ARF and CDKN2B in bladder cancer by real-time quantitative PCR and multiplex ligation-dependent probe amplification.
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[http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=OMIM&cmd=Limits]
Lot 1008: As compared to the previous versions(lot 0107 and 0606), four extra control fragments at 88-96-100-105 nt have been added. This SALSA MLPA kit provides an easy to perform method for the detection of copy number changes of genes located in CDKN2A-CDKN2B (p14ARF-p15-p16) region at 9p21. This region is of interest for several reasons. Firstly, in a significant percentage of various tumour types, both copies of the CDKN2A (p16) gene are inactivated, often because of chromosomal deletions. Secondly, the MTAP gene - located at a only 110 Kb from the CDKN2A gene - encodes for methylthioadenosine phosphorylase, an important enzyme for the salvage of both adenine and methionine. For some time, it has been known that many tumour cells require addition of methionine to their growth medium because their MTAP gene is co-deleted with CDKN2A. Cells lacking MTAP are expected to be sensitive to purine synthesis inhibitors and/or methionine starvation. Germ-line mutations in the CDKN2A gene have been linked to development of cutaneous melanoma in some families with hereditary melanoma. Up to 40% of familial melanoma predisposition is associated with CDKN2A mutations (Hum. Mol. Genet. (2002) 11: 1273-1279). Among the germ-line mutations identified in this gene are several deletions. Disruption of the p14-ARF gene that overlaps with the CDKN2A gene might be the critical abnormality of the melanoma-astrocytoma syndrome which is characterised by a dual predisposition to melanoma and neural system tumours, often astrocytomas (Hum. Mol. Genet. 2001) 10, 55-62. This P024-B probemix contains 13 different probes for the CDKN2A-CDKN2B genes. In addition, it contains three probes for the MTAP gene, 8 probes surrounding these genes in the 9p21 region, as well as 15 reference probes for sequences located in other genes. This SALSA MLPA kit is designed to detect deletions/duplications of one or more exons of the CDKN2A and CDKN2B genes. Heterozygote deletions of probe recognition sequences should give a 35-50% reduced relative peak area of the amplification product of that probe. However, mutations and/or polymorphisms very close to the probe ligation site may also result in a reduced relative peak area. Therefore, apparent deletions detected by a single probe 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.
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