Description
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 (Herman, 1994; Merlo, 1995). Glutathione S-Transferase (GSTpi) 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.
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 GSTpi Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.
The components of the CpG WIZ GSTpi Amplification Kit include those required for PCR amplification after bisulfite modification of DNA samples. Sufficient reagents are provided to analyze 25 samples.
Other Notes
The components of the CpG WIZ GSTpi 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 (neutral cap) 90522 -15°C to -25°C
M Primer Set7.5 μM each primer (25X) 35 μL (red cap) 90523 -15°C to -25°C
W Primer Set7.5 μM each primer (25X) 35 μL (green cap) 90524 -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
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
CPGWIZ is a trademark of Merck KGaA, Darmstadt, Germany
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.
