SALSA MLPA P419 CDKN2A/2B-CDK4 probemix

application: Familial melanoma
region: CDKN2A, CDKN2B, CDK4, MITF E318K
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version: A2
sold since: 2015-08-19

item no. description price
P419-025R SALSA MLPA P419 CDKN2A/2B-CDK4 probemix – 25 rxn € 237
P419-050R SALSA MLPA P419 CDKN2A/2B-CDK4 probemix – 50 rxn € 474
P419-100R SALSA MLPA P419 CDKN2A/2B-CDK4 probemix – 100 rxn € 948
EK1-FAM SALSA MLPA EK1 reagent kit – 100 rxn - FAM € 294
EK1-Cy5 SALSA MLPA EK1 reagent kit – 100 rxn - Cy5 € 294
EK5-FAM SALSA MLPA EK5 reagent kit – 500 rxn - FAM € 1355
EK5-Cy5 SALSA MLPA EK5 reagent kit – 500 rxn - Cy5 € 1355

Please note that both a probemix and reagent kit are needed to perform MLPA.

Malignant melanoma is estimated to be hereditary in 5-10% of the cases. Familial cutaneous melanoma arises in an autosomal-dominant pattern within the affected families and germline alterations in CDKN2A gene are detected in up to 40% of cases (Goldstein AM et al. 2007, J Med Genet. 44:99-106). The CDKN2A gene encodes for two proteins; p16INK4A and p14ARF. The majority of CDKN2A mutations detected in familial melanoma patients affect the exons 2 and 3, which are coding for the p16INK4A. However, exon 1 (previously known as 1β) mutations specific for p14ARF are also reported (Hewitt C et al. 2002, Hum Mol Gen. 11:1273-9), as well as larger genomic deletions of CDKN2A (Randerson-Moor JA et al. 2001, Hum Mol Gen. 10:55-62; Knappskog S et al. 2006, Genes Chromosomes Cancer. 45:1155-63; Laud K et al. 2006, J Med Genet. 43:39-47; Lesueur F et al. 2008, Br J Cancer. 99:364-70). Moreover, homozygous deletions harbouring both CDKN2A and CDKN2B are frequently detected in somatic melanoma samples (Flores JF et al. 1996, Cancer Res. 56:5023-32; Walker GJ et al. 1998, Genes Chromosomes Cancer. 22:157-63).

Another high-penetrance, but low-frequency melanoma susceptibility gene is CDK4, which is mutated 2% of the melanoma families (Goldstein AM et al. 2007, J Med Genet. 44:99-106). Two point mutations (R24H and R24K) of CDK4 have been detected in familial melanoma (Zuo L et al. 1996, Nat Genet. 12:97-9; Soufir N et al. 1998, Hum Mol Genet. 7:209-16; Molven A et al. 2005, Genes Chromosomes Cancer. 44:10-8). Recently, also a germline mutation E318K (952G>A) of MITF gene has been suggested to associate with predisposition to familial melanoma (Yokoyama S et al. 2011, Nature. 480:99-103; Bertolotto C et al. 2011, Nature. 480:94-8).

This probemix contains 14 probes for CDKN2A gene (at 9p21.3), eight probes for CDKN2B gene (at 9p21.3), nine probes for CDK4 gene (at 12q14.1) including a WT probe for CDK4 codon 24, and in addition, 10 probes in the flanking regions of chromosome 9. Furthermore, this probemix also contains a mutation-specific probe for the MITF E318K (952G>A) point mutation. Finally, 14 reference probes have been included, detecting different autosomal chromosomal locations, which are expected to be relatively stable by copy number in melanocytic tumours. This probemix can be used both on germline DNA and on tumour DNA. However, it should be noticed that melanoma karyotypes can harbour multiple numerical and structural aberrations, which can complicate interpretation of the reference probes included in this probemix.

SD008 Sample DNA
Please note that the mutation-specific probe has only been tested on control plasmids and not on positive human DNA samples with the MITF E318K (952G>A) point mutation! This SD008 sample DNA is provided with each probemix vial and can be used in data binning in the fragment analysis and as a positive control for the mutation-specific probe (see next page).

This SALSA® MLPA® probemix is designed to detect copy number changes of one or more sequences in the above mentioned genes and/or chromosomal regions as well as identify the presence of the aforementioned mentioned point mutation in a DNA sample. Heterozygous deletions of recognition sequences should give a 35-50% reduced relative peak height of the amplification product of that probe. Note that a mutation or polymorphism (e.g. SNP) in the sequence detected by a probe can also cause a reduction in relative peak height, even when not located exactly on the ligation site! In addition, some probe signals are more sensitive to sample purity and small changes in experimental conditions. Therefore, single probe deletions/duplications detected by MLPA should always be confirmed by other methods or by MLPA probemixes with higher resolution in the gene or chromosomal area of interest. Users should always verify the latest scientific literature when interpreting their findings. We have no information on what percentage of defects in these genes is caused by deletions/duplications of complete exons. Finally, note that most defects in this gene are expected to be small (point) mutations which will not be detected by this SALSA® MLPA® test.

product history
version A2: As compared to previous lot A1-0412, several probes have a small change in length, but no change in sequence detected.
version A1: changes not specified

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