詳細(xì)說明
| Replacement Information |
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| Description | |
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| Catalogue Number | S7803 |
| Brand Family | Chemicon? |
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| Description | CpG WIZ? MGMT - Methylation specific PCR assay |
| Overview | 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. |
| Materials Required but Not Delivered | Equipment and Supplies a. Microcentrifuge tubes for PCR amplification b. Aerosol-resistant pipette tips c. Thermocycler d. Gel electrophoresis apparatus (vertical or horizontal) e. Power Supply f. 302 nm UV transilluminator, camera and film Reagents a 2.5 mM dNTP mix (2.5 mM of each nucleotide) b. "Hot start" Taq polymerase c. "Hot start" PCR reagents (see Sec. II. Protocols). d. Reagents for gel electrophoresis (1X TBE and 2% agarose, 10% acrylamide, or suitable high resolution agarose) e. DNA markers (size range 100-300 bp) f. Ethidium bromide (10 mg/mL) g. Gel-loading solution / Loading Dye h. Bisulfite Modified DNA (CpGenome? DNA Modification Kit, S7820) |
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| Application | 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. |
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| Application Notes | 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. |
| Biological Information | |
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| UniProt Summary | FUNCTION: SwissProt: P16455 # Involved in the cellular defense against the biological effects of O6-methylguanine (O6-MeG) in DNA. Repairs alkylated guanine in DNA by stoichiometrically transferring the alkyl group at the O-6 position to a cysteine residue in the enzyme. This is a suicide reaction: the enzyme is irreversibly inactivated. COFACTOR: Binds 1 zinc ion. SIZE: 207 amino acids; 21646 Da SUBCELLULAR LOCATION: Nucleus. SIMILARITY: SwissProt: P16455 ## Belongs to the MGMT family. |
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| Material Size | 25 assays |
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