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EpiMark® Bisulfite Conversion Kit

Catalog # Concentration Size List Price Quantity Your Price
E3318S 48 reactions
Please Inquire
Catalog # Size List Price Your Price
E3318S 48 reactions
Please Inquire
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Bisulfite conversion is the most commonly used technique to date and is the “gold standard” for methylation analysis.

  • Incubation of the target DNA with sodium bisulfite results in conversion of all unmodified cytosines to uracils leaving the modified bases (5-mC or 5-hmC) intact
  • Complete conversion of Cytosine is achieved by alternating cycles of thermal denaturation with incubation reactions
  • This kit provides materials for 48 reactions
Genomic DNA from many organisms has modified nucleotides. In the mammalian genome, the modified base is predominately 5-methylcytosine (5-mC), which is involved in gene expression regulation, including selective inactivation of one X chromosome in females of mammalian species (1,2). Symmetrical CpG DNA methylation is heritable, but also reversible. This change of methylation status results in an enormous number of combinations of epigenetic states that can regulate gene expression. In mammalian cells, DNA methylation mainly occurs in CpG dinucleotides and is carried out by two methyltransferase enzymatic activities, namely, maintenance methylation and de novo methylation. The maintenance methyltransferase, DNMT1, is involved in the DNA methylation after every cellular DNA replication cycle. DNMT3a and DNMT3b are the de novo methyltransferases that are active in early development.

For optimal results, 50 ng–2 μg DNA is recommended.

Kit Components: 
Each EpiMark Bisulfite Conversion Kit contains sufficient reagents to perform the bisulfite conversion reaction of 48 samples. Store at room temperature (15–20°C). However, for long-term storage (longer than 4 weeks) Desulphonation Reaction Buffer concentrate should be stored at 2–8°C. Prepared Bisulfite Mix can be stored at -20°C for up to six months. 

Properties & Usage

Materials Required but not Supplied

Custom design primers
EpiMark Hot Start Taq DNA Polymerase (NEB #M0490) or other hot start DNA polymerase specific for bisulfite converted DNA.
Heat block or water bath (requiring temperatures of 65°C and 92°C)
96–100% molecular biology grade ethanol
PCR Thermal Cycler
0.2 ml strip tubes and caps
1.5 ml reaction tubes
Nuclease-free Water
Pipettors and pipette tips (for minimization of cross contamination, use aerosol
barrier tips)

Companion Products
References
  • Bestor, T.H., Verdine, G.L. (1994). Curr. Opin. Cell Biol. . 6, 380-389.
  • Ooi, S.K., O'Donnell A.H., Bestor T.H. (2009). J. Cell Sci. 122, 2787-2791.
  • Frommer, M., et. al. (1992). Proc. Natl. Acad. Sci. USA. 89, 1827-1831.
  • Kumaki, Y., Oda, M. and Okano, M. (2008). Nucleic Acids Res. . 36,
Citations
Additional Citations
  • Fröhlich LF, Mrakovcic M, Smole C, Lahiri P, Zatloukal K (2014) Epigenetic silencing of apoptosis-inducing gene expression can be efficiently overcome by combined SAHA and TRAIL treatment in uterine sarcoma cells PLoS One 9(3), e91558.PubMedID: 24618889, DOI: 10.1371/journal.pone.0091558
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Legal And Disclaimer

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.Methylation-specific PCR may be covered by one or more of U.S. Patent Nos. 5,786,146; 6,017,704; 6,200,756 and 6,265,171 and patents based on foreign counterpart applications. No license or rights under these patents to perform methylation-specific PCR is conveyed expressly or by implication to the purchaser of this product. User's of New England Biolabs' products should determine whether they have all the appropriate licenses in place. Further, no warranty is provided that the use of these products will not infringe on the patents referred to above.

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