CpGenome Fast DNA Modification Kit

Methylation of cytosines located 5′ to guanosine is known to have a profound effect on the expression of many eukaryotic genes. In normal cells methylation occurs predominantly in CG-poor regions, while CG-rich areas, called CpG-islands remain unmethylated. The exceptions are the extensive methylation of CpG islands associated with transcriptional inactivation of regulatory regions of imprinted genes and genes on the inactive X-chromosome of females. Aberrant methylation of normally unmethylated CpG islands has been documented as a relatively frequent event in immortalized and transformed cells and has been associated with transcriptional inactivation of defined tumor suppresser genes in human cancers (7, 8). Hundreds of CpG islands are now known to exhibit the characteristic of hypermethylation in tumors.

Several methods have been developed to determine the methylation status of cytosine. These include digestion with methylation sensitive restriction enzymes as in restriction landmark genomic scanning, oligonucleotide arrays, bisulfite genomic DNA sequencing and Methylation Specific PCR (MSP). Some techniques are more useful for discovery while others are better used for monitoring of known methylated cytosines. Genomic DNA sequencing, although time consuming and labor intensive, offers a more universal detection method. MSP is now an established technology for the monitoring of abnormal gene methylation in selected gene sequences. Utilizing small amounts of DNA, this procedure offers sensitive and specific detection of 5-methylcytosine in promoters. It is being exploited to define tumor suppresser gene function, and to provide a new strategy for early tumor detection.

The initial step of both bisulfite genomic sequencing and MSP is to perform a bisulfite modification of the DNA sample. MSP then involves PCR amplification with specific primers designed to distinguish methylated from unmethylated DNA. The CpGenome Fast DNA Modification Kit contains the reagents for the initial bisulfite modification of the DNA required for both methodologies.

The CpGenome Fast DNA Modification Kit contains reagents required to perform a bisulfite modification on a DNA sample including DNA isolation columns for easy and efficient recovery of bisulfite treated DNA.

For MSP primer design, please use the MethPrime software package. Click here

The CpGenome Fast DNA Modification Kit contains the reagents for the initial bisulfite modification of the DNA required for both methodologies.

25 samples

90584 DNA Modification Reagent 6 g Room Temp

90585 Binding Buffer 20 mL Room Temp

90586 Wash Buffer* 12 mL Room Temp

90587 Elution Buffer 1.5 mL Room Temp

90588 DNA Modification Columns 25 Room Temp

Sufficient reagents are provided in the CpGenome Fast DNA Modification Kit to perform 25 bisulfite reactions.

* Wash Buffer requires the addition of 100% Ethanol.

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