产品说明
一般描述
MSP, performed using the CpGenome
Methylation-specific PCR (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
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, 4). The retinoblastoma 1 (RB1) gene exhibits characteristic hypermethylation im many maligninant tumors.
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 (5). 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
应用
MSP, performed using the CpGenome DNA Modification Kit & the CpG WIZ RB1 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 (neutral cap) 90552 -15°C to -25°C
M Primer Set7.5 μM each primer (25X) 35 μL (red cap) 90553 -15°C to -25°C
W Primer Set7.5 μM each primer (25X) 35 μL (green cap) 90554 -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
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_000321.2 |
UniProt登记号 | P06400 |
application(s) | genomic analysis |
Gene Information | human ... RB1(5925) |
安全信息
储存分类代码 | 10 - Combustible liquids |
Sigma-Aldrich