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Developed Speci f ica l ly for Epigenet ic Research

5-Hydroxymethylcytosine (5hmC) is a modified DNA base (Fig. 1) recently identified as present in mammalian genomes. Referred to as the sixth base, 5hmC is the oxidative product of 5-methylcytosine catalysed by the Ten-eleven translocation (TET) family of enzymes. 5hmC has important roles in regulation of gene expression during normal embryonic development, stem cell reprogramming and disease pathogenesis.

With many years experience and know-how in the manufacture of molecular biology grade nucleotides, Bioline has developed a novel method for producing highly purified 5-hydroxymethyltriphosphate (Hydroxymethyl dCTP). This powerful tool has opened up multiple avenues of investigation for epigenetics research.

Features

• U l t ra-h igh pur i t y >99% t r iphosphate by HPLC

• Readi ly incorporated by standard DNA polymerases

• Hydrox ymethy lated subst rates can be l igated by standard l igases

• DNase, RNase and Nickase f ree

Appl icat ions

• DNA methy lat ion research

• Studies of hydrox ymethy lated DNA /prote in interact ion

• S i te-d i rected mutagenesis

• Subst i tut ion of dCTP in a wide var iet y of molecular b io logy assays

• Label l ing of DNA in v i t ro

EPIK™ Hydroxymethyl dCTP

Bringing you the Sixth Base

United Kingdom Tel: +44 (0)20 8830 5300GermanyTel: +49 (0)3371 681 229AustraliaTel: +61 (0)2 9209 4180

USAToll Free: 888 257 5155FranceTel: +33 (0)1 42 56 04 40SingaporeTel: 1800 BIOLINE

www.bioline.com/epik

Ordering InformationPRODUCT PACK SIZE PRESENTATION CAT NO.

EPIKTM HMdCTP 25μmol 100mM (1 x 250μl) BIO-39046

RELATED PRODUCTS

EPIKTM Amplification Kit 200 Reactions 4 x 1.25ml BIO-66025

dATP 25μmol 100mM (1 x 250μl) BIO-39036

dCTP 25μmol 100mM (1 x 250μl) BIO-39038

dGTP 25μmol 100mM (1 x 250μl) BIO-39037

dTTP 25μmol 100mM (1 x 250μl) BIO-39039

dUTP 25μmol 100mM (1 x 250μl) BIO-39035

EPIK™ Hydroxymethyl dCTP can be used in various molecular biological applications including PCR, where it is readily incorporated by DNA polymerases such as the EPIK™ Amplification Kit from Bioline. This ability to generate PCR products where cytosine is uniformly replaced by hydroxymethylated cytosine can be applied to: epigenetic/methylation studies, studies of hydroxymethylated DNA/protein interaction and forensic DNA analysis. Further applications include the production of DNA fragments resistant to cleavage by restriction endonucleases and generation of cDNA libraries.

EPIK™ Hydroxymethyl dCTP has the distinct advantage of being supplied as lithium salt in highly purified water to ensure functionality and long-term stability. Lithium salt has greater resistance to repeated freeze-thaw cycles than sodium salts and lithium salt preparations remain sterile over the entire shelf-life due to the bacteriostatic activity of lithium towards various microorganisms.

For more information regarding Bioline’s Bulk, Custom and OEM nucleotide service please visit our dedicated microsite: www.bioline.com/nucleotides.

For further details of the new EPIK™ range of epigenetic tools, please visit: www.bioline.com/epigenetics

EPIK™ Hydroxymethyl dCTP

Product Citations1. Nestor, C.E., & Meehan, R.R. Func. Anal. DNA and Chromatin. 259-267 (2014).2. Zhou, T. et al. Mol. Cell 54(5), 879-886 (2014).3. Huh, Y.H. et al., PNAS 110(42), 16862-16867 (2013).4. Pastor, W.A., et al. Nature Protocols 7(10), 1909-1917 (2012).5. Guo, J.U., et al. Cell 145(3), 423-434 (2011).6. Stroud, H., et al. Genome Biol. 12(6), R54 (2011).7. Kinney, S.M., et al. J. Biol. Chem. 286, 24685-24693 (2011).8. Robertson, A.B., et al. NAR 39, e55 (2011).9. Huang, Y., et al. PLoS ONE 5(1), e8888 (2010).10. Tahiliani, M., et al. Science 324, 930-935 (2009).

Physical constants, spectral and HPLC analysis

Product EPIK™ HMdCTP Lithium 100mM Solution

Nomenclature 5 hydroxymethyl 2´-deoxycytidine-5´-triphosphate

Formula C11H14N3O14P3Li4Molecular Weight 520.918g/mol

lmax pH 7.0 275 ± 1nm

e at lmax, pH 7.0 7700 M-1 x cm-1

A250/A260 0.89 ± 0.03

A280/A260 1.27 ± 0.03

Concentration 100mM ± 5%

Appearance Clear Colorless Solution

pH of Solution 7.5 – 8.0

Purity ≥ 99%

DNases, RNases, Nicking Activity

Negative

Storage at –20°C

Stability >24 months

EPIK is a trademark of Bioline Reagents Ltd.

PSGBL0814V1.0

C 5mC 5hmC

Cytosine 5-Methylcytosine 5-Hydroxymethylcytosine

Fig. 1 Cytosine, 5-Methyl and 5-Hydroxymethyl variants