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for Recombinant PvuRts1 I (PvuRts1I) restriction enzymeContents
50 Units PvuRts1 I restriction enzyme
10X PvuRts1 I Reaction Buffer
1 M DTT
for Recombinant PvuRts1 I (PvuRts1I) restriction enzymeBackground
In mammalian genomes, DNA methylation usually occurs at the fifth carbon of cytosine residues. The recent discovery that the TET family of iron-dependent deoxygenases can convert 5-methylcytosine (5-mC) into 5-hydroxymethylcytosine (5-hmC) has raised questions about the functional relevance of 5-hmC in mammalian genomes. The PvuRts1 I restriction enzyme serves as a valuable tool to analyze 5-hmC patterns within the genome. PvuRts1 I is able not only to distinguish between 5-mC and 5-hmC DNA, but it can also cleave both glucosylated and non-glucosylated 5-hmC DNA.
for Recombinant PvuRts1 I (PvuRts1I) restriction enzymeApplication Notes
PvuRts1 I restriction enzyme can be used to directly cleave hydroxymethylated DNA in its non-glucosylated form. The PvuRts1 I enzyme also digests α and β-glucosylated hydroxymethylcytosine DNA, but at a lower efficiency than the non-glucosylated form. The enzyme is specific to 5-hmC DNA and will not digest 5-methylcytosine residues or unmethylated DNA. The enzyme is best suited for use in regions of high 5-hmC density, such as gene bodies.
for Recombinant PvuRts1 I (PvuRts1I) restriction enzymeProtein Details
PvuRts1 I restriction enzyme recognizes the sequence hmCN11-12/N9-10G.
One unit is defined as the amount of enzyme required to fully cleave 1 µg of 5-hmC DNA Standard (Catalog No. 55008) in 30 minutes at 22°C in a total reaction volume of 30 µl. A pilot titration of enzyme is recommended for cleavage of genomic DNA.
PvuRts1 I restriction enzyme digestion of 5-hmC DNA shown by agarose gel.
One µg of unmethylated (Un), 5-methylcytosine (mC) or 5-hydroxymethylcytosine (hmC) Methylated DNA Standard (Catalog No. 55008) was incubated in the absence or presence of 1 unit PvuRts1 I enzyme for 30 minutes at 22°C. Each reaction was run on a 2.5% agarose gel alongside a 1 kb DNA ladder.
for Recombinant PvuRts1 I (PvuRts1I) restriction enzymeStorage
Lyophilized proteins can be stored at -20°C or -80°C, preferably desiccated. Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
for Recombinant PvuRts1 I (PvuRts1I) restriction enzymeGuarantee
This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.
