Hydroxy/Methylated Nucleotides

Ready to use 5-Hydroxymethylcytosine mix (dATP, dTTP, dGTP, d5hmCTP) and 5-Methylcytosine dNTP mix (dATP, dTTP, dGTP, d5mCTP) is of ultra high purity; >99% trisphosphate by HPLC.
Readily incorporated into PCR amplicons with ZymoTaq™ or other DNA polymerases.
Free of endo-, exodeoxyribonuclease, ribonuclease, phosphatase and nicking activities.

Product Size Catalog # Price Qty
5-Methylcytosine dNTP Mix 10 mM D1030
$62.00
5-Methyl dCTP 10 mM D1035
$72.00
5-Hydroxymethyl dCTP 100 mM D1045
$138.00
5-Hydroxymethylcytosine dNTP Mix 10 mM D1040
$62.00

About Hydroxy/Methylated Nucleotides

5-(Hydroxy)methylcytosine dNTP Mix and 5-(Hydroxy)methyl dCTP from Zymo Research are of ultra high purity and can be used to generate DNA by PCR using ZymoTaq™ or other DNA polymerases.

  For 5-hmC For 5-mC
Reagent Volume Final Concentration Volume Final Concentration
2X Reaction Buffer 25 µl 1X 25 µl 1X
dNTP Mix 0.5 µl 0.25 mM each dNTP 0.5 µl 0.25 mM each dNTP
Forward Primer (10 µM) Variable 0.3 to 1 µM Variable 0.3 to 1 µM
Reverse Primer (10 µM) Variable 0.3 to 1 µM Variable 0.3 to 1 µM
Template DNA Variable < 200 ng/50 µl Variable < 200 ng/50 µl
ZymoTaq™ DNA Polymerase (5 U/µl) 1 µl 5 U/50 µl 0.4 µl <2 U/50 µl
ddH20 to 50 µl - to 50 µl -
Total Volume 50 µl   50 µl  
 
Note: The final concentration of MgCl2 in the reaction (above) is 1.75 mM. If required, scale reaction reagent volumes accordingly to optimize the MgCl2, primer, and/or template concentrations.


Concentration 5-Hydroxymethylcytosine dNTP Mix: 2.5 µmol/250 µl as a [10 mM] solution. Contains [2.5 mM] of each dNTP (dATP, dTTP, dGTP and d5hmCTP).
5-Hydroxymethyl dCTP: 10µmol/100 µl as a [100 mM] solution.
5-Methylcytosine dNTP Mix: 2.5 µmol/250 µl as a [10 mM] solution. Contains [2.5 mM] of each dNTP (dATP, dTTP, dGTP and d5mCTP).
5-Methyl dCTP: 1µmol/100 µl as a [10 mM] solution.
Applications PCR amplicons containing 5-(hydroxy)methylcytosine are useful in a variety of applications such as:
  • The study of 5-(hydroxy)methylcytosine binding/interacting protein
  • Define methods that distinguish between 5-methylcytosine and 5-hydroxymethylcytosine.
  • Storage 5-(Hydroxy)methylcytosine dNTP Mix and 5-(Hydroxy)methyl dCTP are guaranteed for 12 months at -20°C. Avoid multiple freeze/thaw cycles. For long-term storage, aliquoting is recommended with storage at -80°C.

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    Featured Citations

    5-hyxroxymethylcytosine-containing DNA substrates were synthesized from a plasmid by utilizing the 5-hmC dNTP mix during PCR amplification. Further analysis utilizing these DNA substrates helped confirm the 5-hmC dehydroxymethylation activities of DNA methyltransferases Dnmt3a and Dnmt3b.

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