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SALSA MLPA KIT R009 Inflammation mRNA
[http://www.geneclinics.org/profiles/hht/details.html]
The SALSA R009 inflammation mRNA probe mix contains several probes specific for mRNAs that are strongly induced by treatment of blood in vivo, or in vitro by lipopolysacharids (LPS). The total probe mix contains 40 different probes, among which probes for NFKB, MIP1a, MIP1b, IL1B, IL1Ra, TNFR1, NFKBIa and IL8 mRNAs. MLPA analysis of RNA requires a minimum of only 10 ng total RNA. Strong signals of probes specific for abundant mRNAs can be reduced by inclusion of a competitor oligonucleotide containing the same hybridizing sequence as the synthetic probe oligonucleotides, but lacking the PCR primer sequence. In R009, the probe signals of the B2M, IL8 and IL1B mRNAs are deliberately reduced by the inclusion of competitor oligonucleotides. In view of the huge production of the IL8 and IL1B mRNAs upon LPS stimulation, this probe mix has been designed such that the signals of these probes are very low in RNA samples derived from untreated human blood. The R009 probe mix contains 40 different probes. The amplification products of the MLPA reactions can easily be separated by capillary electrophoresis. Different fluorescent labelled PCR primers are available. The most commonly used label for ABI apparatus is FAM.
Full mix description (word)
Full mix description (pdf)
Last change in probe mix content: Lot 1103 (November 2003) Current Lot Number.: Lot 0207
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)
[http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed&cmd=Retrieve&dopt=AbstractPlus&list_uids=17371859&query_hl=38&itool=pubmed_docsum]
References 2007 -- High-Throughput mRNA Profiling Characterizes the Expression of Inflammatory Molecules in Sepsis Caused by Burkholderia pseudomallei. 2006 -- Expression of inflammation-related genes in endothelial cells is not directly affected by microparticles from preeclamptic patients. 2005-- High throughput mRNAprofiling highlights associations between myocardial infarction and aberrant expression of inflammatory molecules in blood cells. 2003-- Treatment with an anti-CD14 monoclonal antibody delays and inhibits lipopolysaccharide-induced gene expression in humans in vivo.
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