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SALSA MLPA KIT P105 Oligodendroglioma-2
[http://www.geneclinics.org/profiles/hht/details.html]
EGFR deletion variant III (EGFRvIII) can be caused by deletions of exons 2-7 of EGFR. This MLPA kit can be used to determine the loss of TP53, PTEN and CDKN2A genes as well as gains in copy number of the EGFR and ERBB2 (Her2-neu) gene in DNA samples obtained from oligodendroglioma and other tumors. This probemix might also allow detection of deletions of EGFR that result in the EGFR deletion variant III. The P088 probemix has been developed for analysis of the 1p and 19q chromosome arms in oligodendroglioma DNA samples. Please note that this probemix was intended for oligodendroglioma research. For copy number analysis of only PTEN exons, the P225 PTEN probemix is available. NEW: Now available: P105B ERBB2 and P105B EGFR Silencers. In case of a high level amplification of the ERBB2 or the EGFR gene, this amplification will be clearly visible but the results of the other probes might be more difficult to analyse. As only a small part of the MLPA ligation reaction is used for the PCR reaction., it is possible to repeat this PCR reaction. In order to save reagents it might be useful to reduce all volumes of the PCR reaction two fold. Addition of 1-2 ?l silencer solution to a 25 ?l PCR reaction will selectively decrease the signals of the two ERBB2 probes or the 11 different EGFR probes. Without performing a completely new MLPA reaction, it is possible to analyse the other probes in samples with high level ERBB2 or EGFR amplification. The silencer solutions are available only upon request. Price is EUR 40,- for a vial of 100 ?l. Please mention P105 ERBB2 and / or P105B EGFR Silencer solution on your order. This MLPA kit is designed to detect deletions / duplications of one or more exons of the EGFR, TP53, PTEN and CDKN2A 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 will therefore always require confirmation by other methods. We have no information on the percentage of defects in these genes that are 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. 10; 14-05-2007 Last change in probe mix content: Lot 0407 (April 2007) Current Lot Number.: Lot 0407
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 -- Multiplex ligation-dependent probe amplification: a diagnostic tool for simultaneous identification of different genetic markers in glial tumors.
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