Having supplied restriction enzymes to the research community for over 40 years, NEB has earned the reputation of being the leader in enzyme technologies. Working continuously to be worth of that distinction, NEB strives to develop enzyme of the highest purity and unparalleled quality.
- 100% activity in CutSmart® Buffer (over 210 enzymes are available in the same buffer) allowing for easier double digests
Specificity to epigenetically-relevant DNA modifications (5-mC and 5-hmC)
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Standard Protocol for Restriction Enzyme Digests
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CutSmart Restriction Enzyme Buffer
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Double Digestion with NEBcloner
FspEI, an EpiMark® validated product, is a modification-dependent endonuclease which recognizes CmC sites and generates a double-stranded DNA break on the 3´ side of the modified cytosine at N12/N16. Recognized cytosine modifications include C5-methylation (5-mC) and C5-hydroxymethylation (5-hmC) (1).
This enzyme is provided with an Enzyme Activator Solution which may be used for efficient digestion by FspEI.
The most common epigenetic modifications found in eukaryotic organisms are methylation marks at CpG or CHG sites. A subset of these modified sites are recognized and cleaved by FspEI.
At fully methylated CpG sites:
5´. . . C mC G G . . . 3´
3´. . . G G mC C . . . 5´
or CHG sites:
5´. . . C mC H G G . . . 3´
3´. . . G G D mC C . . . 5´
H = A or C or T (not G)
D = A or G or T (not C)
FspEI recognizes each hemi-methylated site individually and cleaves bidirectionally to generate 32 base or 31 base fragments, respectively. These fragments contain the central methylated site and have 4-base 5´ overhangs at each end. FspEI does not cleave unmodified DNA.
Product SourceAn E. coli strain that carries the synthetic FspEI gene from Frankia species EAN1pec.
The following reagents are supplied with this product:
Store at (°C) Concentration CutSmart® Buffer -20 10 X Enzyme Activator Solution -20 30 X
- Product Categories:
- Methylation Dependent Restriction Enzymes for Epigenetics,
- Restriction Enzymes for Epigenetic Analysis (EpiMark® Validated),
- Restriction Endonucleases: C-G
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