5'			G ↓	C	T	A	G	C		3'
3'			C	G	A	T	C ↑	G		5'

Thermo Scientific NheI restriction enzyme recognizes G^CTAGC sites and cuts best at 37°C in Tango buffer (isoschizomers: AsuNHI, BmtI). See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes. Note: Also available as a FastDigest enzyme for rapid DNA digestion.

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Features

• Superior quality—stringent quality control and industry leading manufacturing process
• Convenient color-coded Five Buffer System
• Includes universal Tango buffer for double-digestions
• BSA premixed in reaction buffers
• Wide selection of restriction endonuclease specificities

Applications

• Molecular cloning
• Restriction site mapping
• Genotyping
• Southern blotting
• Restriction fragment length polymorphism (RFLP)
• SNP

Note: Low salt, high glycerol (>5%) concentrations, pH > 8.0 or a large excess of enzyme may result in star activity. NheI is inhibited by salt concentrations above 100 mM. Supercoiled plasmids may require up to 10-fold more NheI for complete digestion than linear DNAs (e.g. 10 units are required to cleave 1 µg of pBR322 DNA.) For methylation sensitivity, refer to product specifications.