McrBC is an endonuclease which cleaves DNA containing methylcytosine* on one or both strands. Sites on the DNA recognized by McrBC consist of two half-sites of the form (G/A)mC. These half-sites can be separated by up to 3 kb, but the optimal separation is 55-103 base pairs.
*5-methylcytosine or 5-hydroxymethylcytosine or N4-methylcytosine
- McrBC requires GTP for DNA cleavage
- Supplied with NEBuffer 2, 100X BSA and 100x GTP
McrBC is an endonuclease which cleaves DNA containing methylcytosine* on one or both strands. McrBC will not act upon unmethylated DNA (1). Sites on the DNA recognized by McrBC consist of two half-sites of the form (G/A)mC. These half-sites can be separated by up to 3 kb, but the optimal separation is 55-103 base pairs (2,3). McrBC requires GTP for cleavage, but in the presence of a non-hydrolyzable analog of GTP, the enzyme will bind to methylated DNA specifically, without cleavage (4).
*5-methylcytosine or 5-hydroxymethylcytosine or N4-methylcytosine (5).
Product SourceThe two component proteins are purified separately from E. coli K-12 strains containing plasmids encoding McrB and McrC (1).
The following reagents are supplied with this product:
Store at (°C) Concentration NEBuffer™ 2 -20 10 X Purified BSA -20 10 X Plasmid DNA for McrBC Linearized methylated (20µl) -20 100 μg/ml GTP -20 100 X
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