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. | ||||
Specifications
| 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: 10 μmol/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: 1 μmol/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. |
