NotI(Cat. No.:RE107)

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RE107-U050

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500U

2500U

10KU

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Description

Restriction endonuclease, commonly referred to as restriction enzyme, is a type of nuclease that recognizes specific deoxynucleotide sequences and cleaves the phosphodiester bonds between two deoxyribonucleotides at defined sites on each strand. As a key component of the "restriction-modification system", its primary biological role is to protect the host from foreign DNA invasion. Currently, it is widely used in various fields, including gene mapping and cloning, gene structure studies, DNA sequencing and analysis, and gene synthesis. This product is a protein expressed from the NotI gene cloned from Nocardia otitidis-caviarum, which selectively recognizes and cleaves the GC/GGCCGC site. The enzyme is rigorously controlled for host nuclease contamination, exhibiting high activity and stringent specificity.

Features

This product exhibits stringent specificity, enabling sequence-specific cleavage at its target DNA site. The recognition sequence is as follows:
5'... GC↓GGCCGC...3'
3'...CGCCGG↑CG...5'

Usage

1)Molecular cloning
2)Genotyping
3)Southern blot hybridization
4)Restriction fragment length polymorphism (RFLP) analysis

Storage

-20℃±5℃。

Components

Component	RE107-U050	RE107-U250	RE107-M001
NotI (10U/μl)	50μl	250μl	1ml
10×NotI Reaction Buffer	250μl	1.25ml	5ml

Activity Definition

One unit is defined as the amount of enzyme required to completely digest 1 µg of λDNA in a 50 µL reaction volume at 37°C within 1 hour.

Notes

1)Do not exceed 10% of total reaction volume with enzyme to prevent star activity from excess glycerol.
2)Contaminants (e.g., salt, EDTA, or ethanol) in the DNA substrate may disrupt small-volume reactions.
3)Add the enzyme last to the reaction mixture.
4)Gently mix components by pipetting up/down or "flicking" the tube, and quick-spin in a microcentrifuge—avoid vortexing.
5)Keep the enzyme on ice after removal from –20°C, minimize exposure to temperatures above –20°C during handling.
6)Store 10× NotI Reaction Buffer at –20°C ± 5°C.
7)NotI is insensitive to Dam/Dcm methylation but blocked by CpG methylation overlapping its site.
8)Isoschizomer: CciNI recognizes the same sequence (GC↓GGCCGC) as NotI.