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SALSA MLPA KIT P161-P162 Neuroblastoma
[http://www.geneclinics.org/profiles/hht/details.html]
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More probes are available in P252/P253/P254 Neuroblastoma! The P161/P162 Neuroblastoma probemixes can be used to detect copy number changes of several chromosomal regions that frequently show copy number changes in neuroblastoma tumors. One Salsa P161-P162 Neuroblastoma kit contains 50 MLPA reactions of the P161 probemix and 50 MLPA reactions of the P162 probemix (as well as reagents worth of 100 reactions). Kits with 100 reactions of either the P161 or P162 probemix are also available upon 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. An more detailed description of the probes contained in P161/P162 can be found on the next page. 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 P161 probemix also contain a vial with P161 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. 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. An explanation of the procedure can be found on page 3, while some examples of the results are shown on page 8. This MLPA kit is designed to detect deletions / 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 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 (word)
Full mix description (pdf)
Last Update Probe mix description: Vs. 05; 20-10-2006 Last change in probe mix content: P161: lot 1105, P162 lot 1105 (November 2005) Current Lot Number.: P161: lot 0606, P162 lot 0606
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 DNA copy number changes at 8q11- 24 in metastasized colorectal cancer. Poster Katrina Gill et al. 2006. MLPA analysis in neuroblastoma: A pilot study of six cell lines.
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