Bringing you the Sixth Base
Hydroxymethyl dCTP
Features
• U l t r a - h i g h p u r i t y > 9 9% t r i p h o s p h a t e b y H P LC
• R e a d i l y i n c o r p o r a t e d b y s t a n d a r d D N A p o l y m e r a s e s
• H y d r ox y m e t h y l a t e d s u b s t r a t e s c a n b e l i g a t e d b y s t a n d a r d l i g a s e s
• D N a s e , R N a s e a n d N i c k a s e f r e e
Applications
• A p o w e r f u l n e w t o o l f o r D N A m e t h y l a t i o n a n d e p i g e n e t i c s r e s e a r c h
• S t u d i e s o f h y d r ox y m e t h y l a t e d D N A /p r o t e i n i n t e r a c t i o n
• S i t e - D i r e c t e d M u t a g e n e s i s
• S u b s t i t u t i o n o f d C T P i n a w i d e v a r i e t y o f m o l e c u l a r b i o l o g y a s s a y s
• L a b e l i n g o f D N A i n v i t r o
5-hmC (5-Hydroxymethylcytosine) is the hydroxylated and methylated version of cytosine described as the “sixth base” of the
genome.
Bioline has developed a novel enzymatic synthesis method for producing highly purified Hydroxymethyl dCTP (HMdCTP) that
mimics its biological synthesis in T-even bacteriophages. This powerful new tool opens up multiple avenues of investigation for DNA
methylation and epigenetics research. Highly purified HMdCTP (>99% pure by HPLC, Fig.1) can be used in a number of molecular
biological applications.
United Kingdom
Tel: +44 (0)20 8830 5300
USA
Toll Free: 888 257 5155
Germany
Tel: +49 (0)3371 681 229
Australia
Tel: +61 (0)2 9209 4180
www.bioline.com/nucleotides
Hydroxymethyl dCTP
HMdCTP can be used as a substrate for PCR amplification of DNA containing hydroxymethylated
cytosine in place of cytosine. HMdCTP can be used to discriminate between the different DNA molecules
synthesized in one or several PCR cycles. By the use of appropriate enzymes, it is possible to separate the
unhydroxymethylated starting material from the hemi-hydroxymethylated intermediate (produced by a single
primer extension reaction) and from the fully-hydroxymethylated end product.
This ability to generate PCR products whereby cytosine is uniformly replaced by hydroxymethylated
cytosine can also be applied to: epigenetic/methylation assays, studies of hydroxymethylated DNA/protein
interaction, and forensic DNA analysis. Further applications include developing novel strategies for sitedirected mutagenesis and synthesizing DNA fragments resistant to cleavage by a wide range of restriction
endonucleases as well as generation of cDNA libraries.
For more information regarding Bioline’s Bulk, Custom and OEM nucleotide service please visit our dedicated
microsite.
Physical constants, spectral and HPLC analysis
Product
HMdCTP Lithium 100mM Solution
Nomenclature
5 hydroxymethyl 2´-deoxycytidine-5´-triphosphate
Formula
C11H14N3O14P3Li4
Molecular Weight
520.918g/mol
lmax pH 7.0
275nm
e at lmax
7.7 E x mmol-1 x cm-1
Concentration
100mM±2%
Appearance
Clear Colorless Solution
pH of Solution
7.5
dNTP (HPLC Area)
>99%
dNDP (HPLC Area)
<1%
DNases, RNases,
Nicking Activity
Negative
Storage
at –20°C
Stability
>24 months
A 250/A 260
0.90±0.03
A 250/A 260
1.33±0.03
Storage
at –20°C
Stability
>24 months
Product Citations
1. Guo, J.U., et al. Cell 145(3), 423-434 (2011).
2. Stroud, H., et al. Genome Biol. 12(6), R54 (2011).
3. Kinney, S.M., et al. J. Biol. Chem. 286, 24685-24693 (2011).
4. Robertson, A.B., et al. Nucleic Acids Res. 39, e55 (2011).
5. Szwagierczak, A., et al. Nucleic Acids Res. 39(12), 5149-5156 (2011).
6. Huang, Y., et al. PLoS ONE 5(1), e8888 (2010).
7. Jin, S-G., et al. Nucleic Acids Res. 38, e125 (2010).
8. Szwagierczak, A., et al. Nucleic Acids Res. 38(19), e181 (2010).
9. Nestor, C., et al. Biotechniques 48, 317-319 (2010).
10.Tahiliani, M., et al. Science 324, 930-935 (2009).
C
C
Cytosine
USA
Toll Free: 888 257 5155
Germany
Tel: +49 (0)3371 681 229
Australia
Tel: +61 (0)2 9209 4180
5hm
5-Methylcytosine 5-Hydroxymethylcytosine
Fig. 1 HPLC Profile of >99% pure HMdCTP (Hydroxymethylcytosine)
Ordering Information
PRODUCT
dATP
PACK SIZE
PRESENTATION
CAT NO.
25μmol
100mM (1 x 250μl)
BIO-39036
BIO-39038
dCTP
25μmol
100mM (1 x 250μl)
dGTP
25μmol
100mM (1 x 250μl)
BIO-39037
dTTP
25μmol
100mM (1 x 250μl)
BIO-39039
dUTP
25μmol
100mM (1 x 250μl)
BIO-39035
HMdCTP
25μmol
100mM (1 x 250μl)
BIO-39046
P S G B L 0 312 V1.0
United Kingdom
Tel: +44 (0)20 8830 5300
C
5m
www.bioline.com/nucleotides