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S7800 Datasheet(PDF) 5 Page - List of Unclassifed Manufacturers |
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S7800 Datasheet(HTML) 5 Page - List of Unclassifed Manufacturers |
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5 / 32 page  1 I. INTRODUCTION Using This Manual Please read the entire instruction manual prior to using CpG WIZ™ Amplification Kits. Note that in the Procedures and Troubleshooting sections, instructions are separate for the CpG WIZ™ Prader-Willi/ Angelman Amplification Kit (S7806). Should additional questions arise, assistance is available from Chemicon Technical Service at techserv@chemicon.com or (800) 437-7500. Background Methylation of cytosines located 5' to guanosine is known to have a profound effect on the expression of several eukaryotic genes (1). In normal cells, methylation occurs predominantly in CG-poor regions, while CG-rich areas, called CpG islands, remain unmethylated. The exception is extensive methylation of CpG islands associated with transcriptional inactivation of regulatory regions of imprinted genes (2, 3) such as those associated with Prader-Willi/Angelman Syndrome (4) and genes on the inactive X-chromosome of females (5, 6). Aberrant methylation of normally unmethylated CpG islands has been documented as a relatively frequent event in immortalized and transformed cells (7) and has been associated with transcriptional inactivation of defined tumor suppressor genes in human cancers (8, 9). E-cadherin, p16, and p15 are examples of genes that exhibit characteristic hypermethylation. Previously developed methods to determine the methylation status of cytosine include digestion with methylation sensitive restriction enzymes and genomic DNA sequencing. Both techniques have limitations: restriction enzymes can only detect methylation sites within their recognition sequence and sequencing is time consuming. Increasing the detection sensitivity of CpG island methylation has the potential to define tumor suppressor gene function and provides a new strategy for early tumor detection. Methylation-specific PCR (MSP) is a new technology for sensitive detection of abnormal gene methylation utilizing small amounts of DNA (10). 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. The CpGenome™ DNA Modification Kit (S7820) contains the reagents necessary to perform the initial bisulfite reactions, while CpG WIZ™ Amplification Kits contain the reagents required for the PCR amplification reactions. |
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