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CpG Wiz® MGMT Amplification Kit

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货号：S7803

品牌：Millipore 密理博

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Replacement Information
Description
Catalogue Number S7803
Brand Family Chemicon®
Trade Name
  • CpG Wiz
  • Chemicon
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)
References
Product Information
Components
  • 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
Applications
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.
Key Applications
  • PCR
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
Species Reactivity
  • Human
Entrez Gene Number
Gene Symbol
  • MGMT
  • EC 2.1.1.63
UniProt Number
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.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Usage Statement
  • 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.
Storage and Shipping Information
Packaging Information
Material Size 25 assays
Transport Information
Supplemental Information
Specifications

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