NotI restriction enzyme recognizes the 8-base DNA sequence 5′-GCGGCCGC-3′ and cleaves to generate cohesive 5′ sticky ends with a GGCC overhang. LightNing® NotI is suitable for fast restriction digestion in molecular cloning, plasmid construction, vector preparation, restriction mapping, and analysis of PCR products. As an 8-base cutter, NotI is often useful where a less frequent restriction site is required in cloning or DNA analysis workflows.
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NotI Restriction Enzyme for Fast DNA Digestion
NotI restriction enzyme is used for sequence-specific DNA digestion at GCGGCCGC sites in cloning, plasmid construction, vector preparation and restriction analysis workflows.
- Product: LightNing® NotI
- Recognition sequence: 5′-GCGGCCGC-3′
- Cleavage pattern: 5’…GC↓GGCCGC…3′ / 3’…CGCCGG↑CG…5′
- Sticky end: 5′ GGCC overhang
- Reaction condition: 1× CutOne® Buffer
- Incubation temperature: 37°C
- Digestion time: 5–15 minutes under recommended conditions
- Heat inactivation: 80°C for 20 minutes
- Storage: -20°C
- Supplier methylation note: cleavage may be blocked or impaired when substrate DNA is methylated by CpG methylase
- Star activity note: 3 hours incubation does not show star activity, but longer incubation may result in star activity
LightNing® enzymes are a series of engineered restriction enzymes that are capable of fast DNA digestion. All LightNing® enzymes show superior activity in the universal CutOne® and CutOne® Color Buffer, and are able to digest DNA in 5~15 minutes. This enables any combination of restriction enzymes to work simultaneously in one reaction tube and eliminates the need for sequential digestions. LightNing® enzymes have passed multiple strict quality controls, and can be used to digest plasmid, genomic and viral DNA as well as PCR products.
CutOne® Color Buffer includes a density reagent along with red and yellow tracking dyes that allow for direct loading of the reaction mixtures on a gel. The red dye of the CutOne® Color Buffer migrates with 2.5 kb double-strand DNA fragments in a 1% agarose gel, and the yellow dye migrates with 10 bp double-strand DNA fragments in a 1% agarose gel.
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