CpG WIZ® RB1 Amplification Kit
from
EMD Millipore
Description
CpG WIZ® RB1 Amplification Kit, MSP, performed using the CpGenome™ DNA Modification Kit and the CpG WIZ® RB1 Amplification Kit, permits sensitive detection of altered DNA. Because this is a PCR-based assay, it is extremely sensitive, facilitating the detection of low numbers of methylated alleles and the study of samples containing small amounts of DNA. MSP also allows examination of all CpG sites, not just those within sequences recognized by methylation sensitive restriction enzymes. Increasing the number of such sites which can be assessed allows rapid, fine mapping of methylation patterns throughout CpG regions. In addition, the bisulfite modification is ideally suited for analysis of CpG islands since it converts the majority of cytosines to uracils, making a region of the genome which is CG-rich less difficult to amplify by PCR. 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® RB1 Amplification Kit are designed to specifically amplify each of the sequences based upon these chemically-induced differences. If the sample DNA was originally unmethylated, a product will be generated after PCR using the U primer set. Conversely, a product will be generated using the M primer set if the sample was originally methylated.
Specifications/Features
The components of the CpG WIZ® RB1 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 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