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Application

Principles of the Technique

Use of either the CpGenome™ DNA Modification Kit (Cat. No. S7820) or the CpGENOME™ Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the CpG WIZ™ RARbeta2 Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.

The components of the CpG WIZ RARbeta2 Amplification Kit include those required for PCR amplification after bisulfite modification of DNA samples. Sufficient reagents are provided to analyze 25 samples.

Components

The components of the CpG WIZ™ RARbeta2 Amplification Kit include those required for PCR amplification after bisulfite modification of DNA samples. Sufficient reagents are provided to analyze 25 samples with appropriate controls.U Primer Set7.5 µM each primer (25X) 35 µL (clear cap) 90595 -15°C to -25°CM Primer Set7.5 µM each primer (25X) 35 µL (red cap) 90596 -15°C to -25°CW Primer Set7.5 µM each primer (25X) 35 µL (green cap) 90597 -15°C to -25°CU control DNA0.1 µg/µL 50 µL (clear cap) 90393 -15°C to -25°CM control DNA0.1 µg/µL 50 µL (red cap) 90394 -15°C to -25°CW control DNA0.05 µg/µL 50 µL (green cap) 90395 -15°C to -25°CUniversal 10X PCR Buffer 265 µL (blue cap) 90396 -15°C to -25°C

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Methylation of cytosines located 5′ to guanosine is known to have a profound effect on the expression of several eukaryotic genes (Bird, 1992). In normal cells, methylation occurs predominantly in CG-poor regions, while CG-rich areas, called CpG islands, remain unmethylated. Aberrant methylation of normally unmethylated CpG islands has been documented as a relatively frequent event in immortalized and transformed cells (Antequera, 1990) and has been associated with transcriptional inactivation of defined tumor suppressor genes in human cancers (Merlo, 1995). The Retinoic Acid Receptor beta2 is expressed at reduced levels in many tumor types as a result of hypermethylation of its promoter region.



Methylation-specific PCR (MSP) is a new technology for sensitive detection of abnormal gene methylation utilizing small amounts of DNA (Herman, 1996). This process employs an initial bisulfite reaction to modify the DNA, followed by PCR amplification with specific primers designed to distinguish methylated from unmethylated DNA.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

CPGWIZ is a trademark of Merck KGaA, Darmstadt, Germany