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SALSA MLPA KIT P251/P252/P253 Neuroblastoma
[http://www.geneclinics.org/profiles/hht/details.html]
The P251/P252/P253 Neuroblastoma probemixes can be used to detect copy number changes of several chromosomal regions that frequently show copy number changes in neuroblastoma tumors. The older neuroblastoma P161-P162 probemixes are still available on request. Neuroblastoma is a relatively common cancer of childhood that usually occurs sporadically (i.e. non-hereditary). Neuroblastoma is characterized by striking clinical heterogeneity, including subsets that show spontaneous tumor regression. The disease is lethal in over 40% of cases and is responsible for 15% of all cancer-related deaths in children. Several acquired genetic alterations such as amplification of the MYCN oncogene, deletions of chromosome bands 1p36 and 11q23 and unbalanced gains of 17q regions have been well-characterized and shown to be correlated with tumor behavior, including response to treatment. SALSA MLPA P251 probemix contains probes for 1p36, 3p22/3p21.3, 11q23, P252 for 2p24.1/MYCN, 2q33, 17p13/TP53, 17q and P252 for 4, 7, 9, 12 and 14. More information about these regions can be read on the next pages. This MLPA kit is designed to detect gains and losses of one or more genes / chromosomal areas. 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 a single probe will therefore always require confirmation by other methods. Samples with high levels of MYCN amplification can result in extremely low signals for other probes, making it difficult to analyze the latter. Therefore, all kits containing the P252 probemix also contain a vial with P252 silencer solution. This silencer solution contains oligonucleotides that can be included in the PCR reaction of the MLPA procedure. Adding the silencer specifically reduces the signal of the two MYCN probes, making it possible to examine changes in other genes/chromosomal areas. Since only part of the MLPA ligation reaction is used for the PCR reaction, samples with high level MYCN amplification can be further analysed by simply repeating the PCR reaction with the remainder of the ligation reaction, this time including the silencer solution.
Full mix description (pdf)
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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Last change in probemix content Current lot number
: lot 0508 (May 2008) : lot 0508
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