NIPPON Genetics EUROPE GmbHwww.nippongenetics.eu
www.n-genetics.com

Source: Nocardia otitidis-caviarum

Restriction Enzyme
Not I
Cat.#FG-NotI

Size
500 units

Reaction conditions
1X FastGene® 10X Buffer III, 37℃
1X FastGene® 10X FastCut Buffer , 37℃
Conc.
10 units/μl

Store at -20℃
Supplied with: FastGene® 10X Buffer III(FG-REB3)
FastGene® 10X FastCut Buffer (FG-REBHF)
6X DNA Loading Buffer
Sterile water
Recognition site

FastGene® FastCut Buffer
FastGene® restriction enzyme can cut substrate DNA in 5-15 minwith FastGene® FastCut Buffer.

1X FastGene® 10X Buffer III
50 mM Tris-HCl (pH 7.9 at 25℃)
100 mM NaCl
10 mM MgCl2
100 μg/ml BSA

Unit definition
Half unit is defined as the amount of enzyme required for >50%
digestion of pSK M2 at 37℃ for 1 hr in 50 μl reaction mixtures.

Quality control
For Research Use Only. Not for use in diagnostic procedures.

Dilution buffer

NIPPON C
Genetics EUROPE GmbHFastGene® Diluent
www.nippongenetics.eu
HeatInactivationwww.n-genetics.com®

FastGene Not I can be inactivated at 65℃ for 20 min.

Methylation sensitivity

dam methylation: Not sensitive
dcm methylation: Not sensitiveCpG methylation: Sensitive

Restriction Enzyme
Not I

Prolonged incubation

A minimum
required to digest 1 μgConc.
substrate
Cat.# amount of enzyme
SizeDNAfor 16 hr; 0.25 U.
FG-NotI
500 units
10 units/μl

Relative activity in FastGene® BuffersStore ®
atBuffer-20℃ I:FastGene
0%50%
FastGene® Buffer II:®
100%
FastGene Buffer III:® 10X Buffer III
Suppliedwith:
FastGene(FG-REB3)®
FastGene Buffer IV:0%® 10X FastCut Buffer (FG-REBHF)
FastGene®
100%
FastGene FastCut Buffer:
6X DNA Loading Buffer
Sterile waterNote
Cleavage of mammalian genomic DNA is blocked by CpG
Recognition
sitemethylation.
Supercoiledplasmids may require up to 5-foldmore enzyme for complete digestion than linear DNA. Cutting ofDNA with less than 8 bases on either side of the recognition site
is poor. It is suitable to generate large DNA fragments due tothe rare occurrence of the recognition site.

- Unit definition assay- Overdigestion assay- Endonuclease assay- Extreme pure assay

Rev.180817

Standard reaction condition- Normal protocol
Component

Final Conc.

VolumeSubstrate DNA

1 μg

X μl

10X FastGene® Buffer III

1X5 μlSource: Nocardia otitidis-caviarum
Not I

10 unit

Sterile waterReaction
conditions

1 μl

up to 50 μl® 10X Buffer III, 37℃
1X FastGene→ Incubate
at 37℃ for 1 hr
1X FastGene® 10X FastCut Buffer , 37℃- Fast protocol

FastGene®Component
FastCut Buffer

Final Conc.
VolumeFastGene® restriction enzyme can cut substrate DNA in 5-15 min
Substrate DNA
1 μg
X μl
with FastGene® FastCut Buffer.
®
1X5 μl
10X FastGene FastCut Buffer® 10x Buffer III
1X NotFastGeneI10 unit
1 μl
50 mM Tris-HCl (pH 7.9 at 25℃)
Sterile waterup to 50 μl
100 mM NaCl→ Incubate
10 mM
MgCl2 at 37℃ for 15 min
100 μg/ml BSA※We recommend 5-10 units of enzyme per µg DNA and 1020 units for genomic DNA in a 1 h digest.

Unit definition

Half unit is defined as the amount of enzyme required for >50%
digestion of pSK M2 at 37℃ for 1 hr in 50 μl reaction mixtures.

Quality control
For Research Use Only. Not for use in diagnostic procedures.

- Unit definition assay- Overdigestion assay- Endonuclease assay- Extreme pure assay

Rev.180817