Úvod Reagencie Basic research SALSA MLPA ME042 CIMP probemix - 25 reactions

SALSA MLPA ME042 CIMP probemix - 25 reactions

SALSA MLPA ME042 CIMP probemix - 25 reactions

application: CpG Island Methylator Phenotype
region: Various Detailní informace

Cena s DPH € 286.77
Cena bez DPH € 237.00
 25 react
Dostupnost Na dotaz
Kód produktu ME042-025R

Nejnovější informace o produktu naleznete exklusivně na stránkách výrobce MRC-Holland www.mlpa.com (nové okno)

Detailní popis SALSA MLPA ME042 CIMP probemix - 25 reactions

ME042-025R SALSA MLPA ME042 CIMP probemix – 25 rxn

MRC-Holland has prepared a mixture of female genomic DNA from healthy individuals and a carefully titrated amount of plasmid that contains the target sequence recognised by several probes present in the selected MLPA probemixes. The use of SD027 in MLPA reactions performed with the selected MLPA probemixes will therefore show a duplication of several sequences. This SD027 can be ordered separately.

This SALSA® MLPA® probemix is for basic research! This probemix enables you to detect aberrant methylation of CpG islands around the transcription start site of 8 genes for which an altered methylation status in the CpG Island Methylator Phenotype (CIMP) has been reported in literature. Interpretation of results obtained with this product can be complicated. MRC-Holland cannot provide assistance with data interpretation. Please email interesting findings and suggestions for improvements to info@mlpa.com.

CpG-islands are located in or near the promoter region or other regulatory regions of approximately 50% of human genes. Aberrant methylation of CpG-islands has been shown to be associated with transcriptional inactivation of genes in a wide spectrum of human cancers. In addition, DNA methylation analysis can indicate in some cases from which type of tissue the tumour was derived.

This ME042-C1 CIMP MS-MLPA probemix contains 31 MS-MLPA probes which detect the methylation status of promoter regions of the following 8  genes: CACNA1G, CDKN2A, CRABP1, IGF2, MLH1, NEUROG1, RUNX3 and SOCS1. The promoter regions of these genes are unmethylated in blood-derived DNA of healthy individuals but have been reported to be frequently methylated in CpG Island Methylator Phenotype (CIMP) tumours. The CIMP status of a tumour may be useful for prediction of treatment response and prognosis in colorectal cancer (Walther A et al. 2009, Nat Rev Cancer, 9:489-99). In addition, as recent studies have shown that a BRAF V600E activating mutation is tightly associated with CIMP positivity (Weisenberg DJ et al. 2006, Nature Genet. 38:787-93), a mutation specific probe detecting this mutation is included. This BRAF probe will only generate a signal when the V600E mutation is present in a DNA sample.

The MLPA technique is widely used for the detection of aberrant copy numbers of DNA sequences. For the detection of aberrant methylation, the MLPA technique is combined with digestion of the sample DNA-probe hybrids with the methylation sensitive restriction endonuclease HhaI. In addition to the MS-MLPA probes, 15 reference probes are included which are not affected by HhaI digestion and are relative stable by copy number in colorectal tumours. Besides detecting aberrant methylation, all MS-MLPA probes will give information on copy number changes in the analysed sample. Also, one digestion control probe is included in this probemix indicating whether or not restriction endonuclease digestion in the MS-MLPA reaction was complete.

SD029 Sample DNA: Please note that the mutation-specific probe for the V600E mutation has only been tested on control plasmids and not on V600E mutation-positive human DNA samples! This SD029 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 probes (see page 3).

The MS-MLPA probes in this ME042-C1 probemix detect sequences in promoter regions of tumour suppressor genes that are unmethylated in most blood-derived DNA samples. Upon digestion, the peak signal obtained in unmethylated samples will be very small or absent. In contrast, when tested on in vitro methylated human DNA, these probes do generate a  signal. We have no data showing that methylation detected by a  particular probe indeed influences the corresponding mRNA levels. The MLPA reaction requires as little as 50 ng of human DNA and can be used on a variety of DNA samples, including those derived from paraffin-embedded tissues.

This SALSA® MS-MLPA® probemix can be used to detect aberrant methylation of one or more sequences of the CACNA1G, CDKN2A, CRABP1, IGF2, MLH1, NEUROG1, RUNX3 and SOCS1 genes. Methylation levels can be different for different tissues. If possible, use identically treated test and reference samples (same tissue type and DNA extraction method). This SALSA® MS-MLPA® probemix can be used also detect the presence of the aforementioned mentioned BRAF V600E 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 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, deletions and duplications detected by MLPA should always be confirmed by other methods. Not all methylation changes, and copy number alterations detected by MLPA will be pathogenic; users should always verify the latest scientific literature when interpreting their findings.

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