产品说明
一般描述
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. Aberrant methylation of normally unmethylated CpG islands has been documented as a relatively frequent event in immortalized and transformed cells (2) and has been associated with transcriptional inactivation of defined tumor suppressor genes in human cancers (3). BRCA1 is a tumor suppressor gene in familial breast-ovarian carcinoma syndrome and in sporadic ovarian carcinomas. In normal breast epithelial cells, BRCA1 mRNA levels in tumors appear to be down-regulated by methylation, while BRCA2 shows significant over expression in sporadic breast cancers. Epigenetic hypermethylation of the BRCA1 gene plays an important role when it is expressed at reduced levels in breast and ovarian tumors as a result of hypermethylation of its promoter region (5,6,7).
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 (4). 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
Principles of the Technique:
Methylation-specific PCR (MSP), performed using the CpGenome
MSP employs an initial bisulfite reaction to modify the DNA, followed by a "hot start" PCR amplification with specific primers designed to distinguish methylated DNA from unmethylated DNA. As shown in Figure 1, in the bisulfite reaction, all unmethylated cytosines are converted to uracils while 5-methylcytosines remain unaltered. Thus, the sequence of the treated DNA will differ if the DNA is originally methylated vs. unmethylated. Primers contained in the CpG WIZ
应用
Methylation-specific PCR (MSP), performed using the CpGenome DNA Modification Kit & the CpG WIZ BRCA1 Amplification Kit, permits sensitive detection of altered DNA.
组分
The components of the CpG WIZ
U Primer Set7.5 μM each primer (25X) / 35 μL (clear cap) / 90645 / -15°C to -25°C
M Primer Set7.5 μM each primer (25X) / 35 μL (red cap) / 90646 / -15°C to -25°C
W Primer Set7.5 μM each primer (25X) / 35 μL (green cap) / 90647 / -15°C to -25°C
U control DNA0.1 μg/μL / 50 μL (clear 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
法律信息
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
COPRIME is a registered trademark of Merck KGaA, Darmstadt, Germany
CPGWIZ is a trademark of Merck KGaA, Darmstadt, Germany
免责声明
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.
基本信息
eCl@ss | 32161000 |
产品性质
质量水平 | 100 |
species reactivity | human |
manufacturer/tradename | CPGWIZ™ Chemicon® |
technique(s) | PCR: suitable |
NCBI登记号 | NM_007294.2 NM_007295.2 NM_007296.2 NM_007297.2 NM_007298.2 NM_007299.2 NM_007300.2 NM_007302.2 NM_007303.2 NM_007304.2 NM_007305.2 |
UniProt登记号 | P38398 |
application(s) | genomic analysis |
运输 | dry ice |
Gene Information | human ... BRCA1(672) |
安全信息
储存分类代码 | 10 - Combustible liquids |