Biofargo NheI, GMP Grade Restriction Enzyme (20 U/µL)

Description

Biofargo NheI, GMP Grade is a recombinant restriction endonuclease expressed in Escherichia coli and supplied at 20 U/µL with its matching 10× CutOne® Buffer, GMP Grade. It recognizes the sequence G^CTAGC and completely digests target DNA in 15 minutes to 1 hour at 37 °C.

The enzyme is manufactured and quality-controlled in compliance with GMP specifications, with full traceability of the production process and raw materials. No antibiotics and no animal-derived raw materials or excipients are used anywhere in production. Process-related impurities are tightly controlled, including host cell protein, host cell DNA, non-specific endonuclease, DNase and RNase activity, microbial limits and bacterial endotoxin. According to the manufacturer, the product meets the requirements for raw materials in fields such as vaccine and pharmaceutical production.

Each lot is tested for host cell DNA, host cell protein, microbial limit, bacterial endotoxin and heavy metals using methods from the Chinese Pharmacopoeia (ChP 2025, Volume IV), and for mycoplasma using a LAMP-based detection kit.

GMP Grade: “GMP Grade” describes the manufacturing and quality-control standard of this reagent. It is not a claim of suitability for clinical use; users are responsible for qualifying the material for their own processes.

Research Use Only: This product is intended for laboratory research use only. It is not intended for diagnostic, therapeutic, food, household, or clinical use.

Specifications

Quality Control

Key Features

• GMP-compliant manufacturing: Produced and quality-controlled under GMP specifications with full traceability of the process and raw materials.
• Animal-component-free and antibiotic-free: No animal-derived materials, excipients or antibiotics are used in production.
• Fast, complete digestion: Digests target DNA in 15 minutes to 1 hour at 37 °C.
• Comprehensive impurity control: Each lot is tested for host cell DNA, host cell protein, endotoxin, microbial limit, mycoplasma and heavy metals.
• High purity: ≥95% pure by SDS-PAGE, with no detectable DNase or non-specific nuclease contamination under the specified test conditions.
• Matched GMP-grade buffer: Supplied with 6 × 1 mL of 10× CutOne® Buffer, GMP Grade.
• Heat inactivation: Inactivated by incubation at 80 °C for 20 minutes.

Applications

• Restriction cloning and subcloning
• Plasmid linearization for in vitro transcription (IVT) templates
• Construction of expression vectors for vaccine and biopharmaceutical development
• Restriction mapping and DNA fragment preparation

Instructions for Use

Combine the following components on ice in the order shown:

^a DNA substrates should be free of phenol, chloroform, ethanol, EDTA, detergents and high salt concentrations, which can reduce enzyme activity.

1. Mix gently by pipetting or flicking the tube (do not vortex), then spin down briefly.
2. Incubate at 37 °C for 15 minutes to 1 hour.
3. Inactivate the enzyme by heating at 80 °C for 20 minutes.

Notes:

• The enzyme volume should not exceed 10% of the total reaction volume, to avoid star activity caused by excess glycerol from the enzyme storage buffer.
• Additives in the enzyme storage buffer (e.g. glycerol, salt) and contaminants in the DNA solution (e.g. salt, EDTA, ethanol) inhibit digestion more strongly in smaller reaction volumes.
• Star activity does not occur under the recommended conditions. Extended incubation or high enzyme concentrations may result in star activity.

Number of Recognition Sites in Common DNAs

Methylation Effects on Digestion

Storage and Handling

• Store at −20 ± 5 °C.
• Shelf life is 24 months when stored under the recommended conditions.
• Shipped at ≤0 °C.
• Keep the enzyme on ice during reaction setup and return it to −20 °C promptly after use.
• Use appropriate laboratory protective equipment when handling.

Documents

NheI, GMP Grade – Instruction Manual (GMP505S)

The Safety Data Sheet (SDS) and lot-specific Certificate of Analysis (COA) are available on request. Please contact us with the catalog number and lot number.

FAQ