CpGenome™ Universal DNA Modification Kit

The CpGenome™ DNA Modification Kit is used to create methylation-specific sequence alterations in any DNA sample. Using chemical treatment, the kit converts all unmethylated cytosine residues to uracil. Methylated cytosine residues are unaltered in this process. Therefore, the chemically-treated DNA sequence will differ only if it was originally unmethylated in vivo. After sequence conversion, methylation specific PCR is employed to detect these differences using primers specific for any gene of interest (or the CpG™ Wiz Amplification Kit Primers).

To download the CpGWARE™ Primer Design Software for use with the CpGenome™ Universal DNA Modification Kit.  Please visit www.cpgware.com, the software can be downloaded for free. The "Username" and "password" is "cpgware" for both entered in small case.

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 CpGenome™ Fast DNA Modification Kit

The CpGenome™ Fast DNA Modification Kit contains reagents required to perform a bisulfite modification on a DNA sample.  In the bisulfite reaction, all unmethylated cytosines are deaminated and sulfonated, converting them to uracils, while 5-methylcytosines remain unaltered. Thus, the sequence of the treated DNA will differ depending on whether the DNA is originally methylated or unmethylated. Also, the initially complementary DNA strands will no longer be complementary after cytosine conversion. Primers for use in MSP can be designed to specifically amplify either a bisulfite-sensitive, unmethylated strand or a bisulfite-resistant, methylated strand, based upon these chemically-induced differences. MSP

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 CpGenome™ Universal Unmethylated DNA

Universal Unmethylated DNA Vial A contains 5 micrograms (50μL) of human genomic DNA at a concentration of 0.1 μg/μL. Universal Unmethylated DNA Vial B contains 5 micrograms (50μL), of genomic DNA isolated from a primary human fetal cell line at a concentration of 0.1 μg/μL. Note: The DNA must first be bisulfite modified using (S7820) to use a specific primer set for the unmethylated form of the gene of interest.

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 CpGenome™ Universal Methylated DNA

CpGenome™ Universal Methylated DNA is enzymatically methylated (for all genes) human male genomic DNA.  This product is intended for use with the CpGenome™  DNA Modification Kit or with MS-PCR amplification primers.  It can be used as a methylation-positive control for gene methylation studies.  Each vial contains 10 micrograms (100μL) of human genomic DNA at a concentration of 0.1μg/μL.

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 CpG WIZ® p15 Amplification Kit

Aberrant methylation of the p15 promoter region has been shown to cause inactivation in several gliomas and leukemias. The CpG WIZ® p15 Amplification Kit is used for determining the methylation status of the p15 promoter by methylation-specific PCR (MSP). The kit contains primers targeted to regions of the p15 promoter where the sequences are most divergent after bisulfite treatment. PCR parameters have been identified so that all primer sets in the kit amplify template under the same conditions. Control genomic DNA samples (methylated and unmethylated) for p15* are also included.

Use of either the CpGenome™ DNA Modification Kit (Cat. No. S7820) or the CpGENOME™ Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the The CpG WIZ® p15 Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.

*p15 is covered under U.S. patents 5,994,095, US 6,140,473, 6,180,776, 6,210,949 and 6,218,146.

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 CpG WIZ® p16 Amplification Kit

Expression of p16*, a tumor suppressor gene, is repressed in a variety of cancers such as bladder, colon and retinoblastoma. Methylation of CpG islands in the p16 promoter has been shown to be responsible for inactivation of this gene in certain cases. The CpG WIZ® p16 Amplification Kit is used for determining the methylation status of the p16 promoter by methylation-specific PCR (MSP). The kit contains primers targeted to regions of the promoter where the sequences are most divergent after bisulfite treatment. PCR parameters have been identified such that all primer sets in the kit amplify under the same conditions. Control genomic DNA samples (methylated and unmethylated) for p16 are also included.
Methylation-specific PCR (MSP) is a new technology for sensitive detection of abnormal gene methylation utilizing small amounts of DNA (Herman, 1996 ). 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.

Use of either the CpGenome™ DNA Modification Kit (Cat. No. S7820) or the CpGENOME™ Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the CpG WIZ® p16 Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.

*p16 is covered under U.S. patents 5,801,236, 5,843,756, and 6,090,578.

Methylation Specific PCR (MSP) of the p16 gene in two invasive carcinomas, a squamous intraepithelial lesion (SIL), and an adenocarcinoma of the cervix. Each numbered set are paired MSP reactions specific for both the unmethylated (U) and methylated (M) alleles of the p16 CpG island. The presence or absence of the "M" MSP amplicon is indicative of the methylation state of the p16 gene in the sample. The results indicate that both invasive carcinomas and the SIL sample are heterozygous for methylation while the adenocarcinoma sample is clearly homozygous for the unmethylated state at the p16 locus.

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 CpG WIZ® Prader-Willi/Angelman Amplification Kit

A differentially methylated site exists in the CpG islands of SNRPN (small nuclear ribonucleoprotein-associated polypeptide N), a candidate gene for Prader-Willi/Angelman Syndrome. Normally, this site is methylated and inactive on the maternal homolog and unmethylated and transcriptionally active on the paternal homolog. In research samples from Prader-Willi individuals, only the methylated allele is present, while samples from Angelman Syndrome (AS) individuals contain only the unmethlyated allele. The CpG WIZ® Prader-Willi/Angelman Amplification Kit is used for determining the methylation status of this region by methylation -specific PCR (MSP). The kit contains primers targeted to the CpG island on the SNRPN gene. PCR parameters have been identified such that both primer sets in the kit amplify template under the same conditions. Normal control genomic DNA, which will generate a product with both primer sets, is also included.


Use of either the CpGenome™ DNA Modification Kit (Cat. No. S7820) or the CpGENOME™ Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the CpG Wiz® Prader-Willi/Angelman Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.

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 CpG WIZ® E-cadherin Amplification Kit

E-cadherin is a cell adhesion protein that suppresses tumor cell invasion and metastasis in tumor models. On mechanism regulating the expression of the E-cadherin gene is methylation of the promoter region. The CpG WIZ® E-cahedrin Amplification Kit is used for determining the methylation of status of the E-cadherin promoter by methylation-specific PCR (MSP). The kit contains primers targeted to regions of the promoter where the sequences are most divergent after bisulfite treatment. PCR parameters have been identified so that all primer sets in the kit amplify under the same conditions. Control genomic DNA samples (methylated and unmethylated) for E-cadherin are also included.

Alternate Names: CD324

Use of either the CpGenome™ DNA Modification Kit (Cat. No. S7820) or the CpGENOME™ Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the CpG WIZ® E-cadherin Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.

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 CpG WIZ® DAP-Kinase Amplification Kit

The components of the CpG WIZ® DAP-kinase 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.

Use of either the CpGenome™ DNA Modification Kit (Cat. No. S7820) or the CpGENOME™ Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the CpG WIZ® DAP-kinase Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.

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 CpG WIZ® RASSF1A Amplification Kit

The components of the CpG WIZ® RASSF1A 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.

Use of either the CpGenome™ DNA Modification Kit (Cat. No. S7820) or the CpGENOME™ Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the CpG WIZ® RASSF1A Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.

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 CpG WIZ® MGMT Amplification Kit

The components of the CpG WIZ® MGMT 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.

Use of either the CpGenome™ DNA Modification Kit (Cat. No. S7820) or the CpGENOME™ Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the CpG WIZ® MGMT Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.

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 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.   by PCR.

The CpGenome™ DNA Modification Kit (S7820) contains the reagents necessary to perform the initial bisulfite reactions, while the CpG WIZ® RB1 Amplification Kit contains the reagents required for the PCR amplification reactions.

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.

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 CpG WIZ® Fragile X Amplification Kit

The components of the CpG WIZ® Fragile X 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.

Use of either the CpGenome™ DNA Modification Kit (Cat. No. S7820) or the CpGENOME™ Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the CpG WIZ® Fragile X Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.

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 CpG WIZ® SOCS1 Amplification Kit

Methylation of cytosines located 5' to guanosine is known to have a profound effect on the expression of several eukaryotic genes (Bird, 1992). 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 (Antequera, 1990) and has been associated with transcriptional inactivation of defined tumor suppressor genes in human cancers (Herman, 1994; Merlo, 1995). The suppressor of cytokine signaling-1 (SOCS1) gene exhibits characteristic hypermethylation.

The components of the CpG WIZ® SOCS1 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.

Use of either the CpGenome™ DNA Modification Kit (Cat. No. S7820) or the CpGENOME™ Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the CpG WIZ® SOCS1 Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.

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 CpG WIZ® hMLH1 Amplification Kit

The components of the CpG WIZ® hMLH1 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.

Use of either the CpGenome™ DNA Modification Kit (Cat. No. S7820) or the CpGENOME™ Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the CpG WIZ® HMLH1 Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.

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 CpG WIZ® VHL Amplification Kit

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.

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.

Principles of the Technique

MSP, performed using the CpGenome™ DNA Modification Kit and the CpG WIZ® VHL Amplification Kit, permits sensitive detection of altered DNA.  Primers contained in the CpG WIZ® VHL Amplification Kit are designed to specifically amplify each of the sequences based upon these chemically-induced sequence 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.

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 CpG WIZ® APC Amplification Kit

The components of the CpG WIZ® APC 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.

Use of either the CpGenome™ DNA Modification Kit (Cat. No. S7820) or the CpGENOME™ Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the The CpG WIZ® APC Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions.

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