CpG WIZ^™ MGMT - Methylation specific PCR assay

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). O(6)-Methylguanine DNA methyltransferase (MGMT) is an example of a gene that exhibits characteristic hypermethylation.



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.

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 MGMT Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.

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

The components of the CpG WIZ MGMT 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 Set5 μM each primer (25X) 35 μL (neutral cap) 90513 -15°C to -25°C

M Primer Set5 μM each primer (25X) 35 μL (red cap) 90514 -15°C to -25°C

W Primer Set5 μM each primer (25X) 35 μL (green cap) 90515 -15°C to -25°C

U control DNA0.1 μg/μL 50 μL (white cap) 90393 -15°C to -25°C

M control DNA0.1 μg/μL 50 μL (red cap) 90394 -15°C to -25°C

W control DNA0.05 μg/μL 50 μL (green cap) 90395 -15°C to -25°C

Universal 10X PCR Buffer 265 μL (blue cap) 90396 -15°C to -25°C

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

CPGWIZ is a trademark of Merck KGaA, Darmstadt, Germany

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