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.
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.
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.