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

Source: Nocardia aerocolonigenes

Restriction Enzyme
Nae I
Cat.#FG-NaeI

Size
500 units

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

Store at -20℃
Supplied with:

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

1X FastGene® Buffer I
10X FastGene® Buffer I
10X FastGene® FastCut Buffer(FG-REB1)(FG-REBHF)

6X DNA Loading BufferSterile water
Recognition site

10 mM Bis Tris propane-HCl (pH 7.0 at 25℃)
10 mM MgCl2
100 μg/ml BSA

Unit definition
One unit is defined as the amount of enzyme required forcomplete digestion of 1 μg pBR322 at 37℃ for 1 hr in 50 μl
reaction mixtures.

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

Dilution buffer
FastGene® Diluent A

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

Heat Inactivation
Nae I can beinactivated
65℃ for20 min.
NIPPONGeneticsatEUROPE
GmbH
www.nippongenetics.eu
Methylation sensitivity
www.n-genetics.com
dam methylation: Not sensitive

dcm methylation: Not sensitive

Standard reaction condition- Normal protocol
Component

Final Conc.
1 μg

X μl

10X FastGene® Buffer I

1X

5 μl

Nae I

4 unit

1 μl

CpG methylation: Sensitive

Source: Nocardia aerocolonigenes

Prolongedincubation
Restriction
EnzymeReactionconditions
Sterile water

Relative activity in FastGene® Buffers

FastGene® FastCut Buffer

A minimum amount of enzyme required to digest 1 μg substrate
DNA for 16 hr; 0.5 U.

Nae ICat.#® Buffer I:
SizeConc.
FastGene100%® Buffer II:
FG-NaeI
500 100%units
4 units/μl
FastGene®
FastGene
25%
Store atBuffer-20℃ III:
100%
FastGene® Buffer IV:®
FastGene FastCut Buffer:100%
Supplied with: 10X FastGene® Buffer I(FG-REB1)
Note
10X FastGene® FastCut Buffer (FG-REBHF)
It is an isoschizomer
NgoM
lV. Cleavage
6XofDNA
Loading
Buffer of mammalian
genomic DNA is blocked
CpG methylation. It displays markedSterile bywatersite preference, while NgoM IV has less site preference. Tworecognitionsequences
Recognitionsite are required for cleavage. One of the twoacts as an effector site. It is sensitively affected by the locationsof the recognition sequence. For example, if the two sites are
too close, Nae I is not efficient in cleaving one of the two.

up to 50 μl

1X FastGene® Buffer I 37℃→ Incubate
at 37℃for 1 37℃
hr
® FastCut
1X FastGene
Buffer,- Fast protocol

Component
FinalConc. DNAVolume
FastGene® restriction
enzyme can cut
substratein 5-15 min® FastCut Buffer.
withSubstrate
FastGeneDNA
1 μg
X μl®

® Buffer
FastGene
FastCutI Buffer
1X 10XFastGene1X

10 mM
Nae BisI Tris propane-HCl (pH 7.0 at425℃)
unit
10 mM MgCl2100Sterileμg/mlwater
BSA→ Incubate at 37℃ for 15 min

5 μl
1 μl
up to 50 μl

Unit definition

Weisrecommend
5-10
units ofofenzymeµg DNA
One ※
unit
defined as theamountenzymeper
required
forand 1020 units
for genomicDNA
in a 1ath37℃
digest.
complete
digestionof 1 μg
pBR322for 1 hr in 50 μl
reaction mixtures.

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

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

VolumeSubstrate DNA

Dilution buffer
FastGene® Diluent A