CpG WIZ® VHL Amplification Kit
from
EMD Millipore
Description
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 von Hippel-Lindau (VHL) gene exhibits 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 (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™ DNA Modification Kit (S7820) contains the reagents necessary to perform the initial bisulfite reactions, while the CpG WIZ® VHL Amplification Kit contains the reagents required for the PCR amplification reactions., CpG WIZ® VHL Amplification Kit
Specifications/Features
The components of the CpG WIZ® VHL 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) 90516 -15°C to -25°C;M Primer Set5 µM each primer (25X) 35 µL (red cap) 90517 -15°C to -25°C;W Primer Set5 µM each primer (25X) 35 µL (green cap) 90518 -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