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Epigenetics |
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Epigenetics>【Zymo】 |
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Hydroxy/Methylated Nucleotides(D1045) |
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Hydroxy/Methylated Nucleotides
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.
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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 |
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50 µl |
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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.
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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. |
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-Methylcytosine dNTP Mix |
10 mM |
D1030 |
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5-Methyl dCTP |
10 mM |
D1035 |
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5-Hydroxymethyl dCTP |
100 mM |
D1045 |
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5-Hydroxymethylcytosine dNTP Mix |
10 mM |
D1040 |
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