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EpiTect II DNA Methylation Enzyme Kit

For preparation of genomic DNA samples for analysis with EpiTect Methyl II PCR Arrays and EpiTect Methyl II PCR Assays using MethylScreen technology

  • Specific methylation-sensitive and methylation-dependent enzymes
  • Optimized conditions for complete DNA digestion
  • For use with EpiTect Methyl II PCR Arrays and Assays

The EpiTect II DNA Methylation Enzyme Kit prepares genomic DNA samples for methylation analysis using real-time PCR primers and arrays for known and predicted methylated CpG islands. The EpiTect II DNA Methylation Enzyme Kit uses MethylScreen technology provided under license from Orion Genomics, LLC.

Cat No./ID: 335452
EpiTect II DNA Methylation Enzyme Kit (12)
S Fr.393.00
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5x Restriction Digestion Buffer, Methylation-Sensitive Enzyme A, Methylation-Dependent Enzyme B
The EpiTect II DNA Methylation Enzyme Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Principle

The EpiTect II DNA Methylation Enzyme Kit prepares genomic DNA samples for DNA methylation analysis using EpiTect Methyl II PCR Assays and EpiTect Methyl II PCR Arrays for individual or panels of known and predicted methylated CpG islands. Using the enzymes and buffer provided in the kit, 4 digests are performed to detect different methylated DNA fractions. The product of a mock digest (Mo) contains all of the input genomic DNA. The product of the methylation-sensitive restriction enzyme mixture (Enzyme A) digest (Ms) contains methylated DNA sequences, while the product of the methylation-dependent restriction enzyme mixture (Enzyme B) digest (Md) contains unmethylated DNA sequences. The product of a double digest (Msd) measures the background and the success of both enzymatic digestions.

Procedure

Simply add equal amounts of a genomic DNA sample to 4 separate tubes into which buffer and the appropriate restriction enzyme combinations are added. The mock digest (Mo) has no enzymes added; Enzyme A is added to the methylation-sensitive restriction digest (Ms); Enzyme B is added to the methylation-dependent restriction digest (Md); both enzyme mixtures are added to the double digest (Msd). All digests are incubated at 37°C for between 6 hours and overnight. Finally, the enzymes are heat inactivated at 65°C for 20 minutes.

For optimal results, the EpiTect II II DNA Methylation Enzyme Kit should be used in conjunction with EpiTect Methyl II PCR Arrays and EpiTect Methyl II PCR Assays to analyze the amount of DNA in each digest of each sample and determine the methylation status of CpG islands in gene promoters or other methylated DNA sequences of interest. Simply dilute each digest into the appropriate RT2 SYBR® Green Mastermix and aliquot into the appropriate well of the array. Or, add an aliquot of each digest to separate real-time PCR tubes containing the appropriate RT2 SYBR® Green Mastermix and the same assay. Finally, run the recommended cycling program.

Please check the kit components immediately after you receive this package. We are not responsible for any missing items not reported within two (2) business days upon receipt.

Tube / Contents  Quantity
5X Restriction Digestion Buffer  660 µl × 4 Tubes
Enzyme A (Methylation-Sensitive)  25 µl in One 0.5-mL Tube
Enzyme B (Methylation-Dependent)  25 µl in One 0.5-mL Tube

Storage Conditions:
The EpiTect Methyl II DNA Restriction Kit (12) is shipped either on dry ice or with blue ice packs and must be stored at -20 ºC. We suggest that all kit contents be kept in their original container to insure that no components are misplaced.

Shelf Life:
All reagents are stable for 6 months after receipt of the kit if stored at the recommended temperature.

Related Products:
EpiTect Methyl II PCR Arrays
EpiTect Methyl II PCR Primer Assays for individual gene promoters
RT² SYBR® Green qPCR Master Mixes (Available for all real-time PCR instruments)

The EpiTect Methyl II DNA Restriction Kit uses the MethylScreen™ Technology provided under license from Orion Genomics, LLC.

Purchaser agrees that use of this product and data therefrom is limited solely to the purchaser and for only the purchaser’s own internal molecular biology research applications (“Permitted Use”), and shall not be re-sold or used for any other purposes (all of which are expressly prohibited), including without limitation diagnostic purposes, uses that could require regulatory approval for diagnostics from an agency of any government or regulatory entity anywhere in the world, diagnosis, prevention, or treatment of disease, and the right to perform commercial services of any kind, including without limitation, reporting the results of purchaser’s activities, including without limitation, for a fee or other commercial consideration. Except for the Permitted Use, no rights, titles, or interests in or to any tangible or intangible property rights are conveyed or shall be deemed conveyed by implication, estoppel or otherwise. The performance characteristics of the product other than for the Permitted Use are unknown.

Applications

The EpiTect II DNA Methylation Enzyme Kit forms part of the EpiTect Methyl II PCR system. This is an innovative technology and versatile tool for:

Methylation pattern profiling
Verification of genomewide methylation analyses
Cancer and stem cell biomarker discovery
Toxicological and epidemiological screening
Characterization of gene expression regulation
Epigenetic DNA methylation analysis

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Kit Handbooks (1)
For pathway- or disease-focused profiling of regional DNA methylation using MethylScreen technology
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