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

Source: E.coli that carries the EcoR V genefrom plasmid J62 pLG74

Restriction Enzyme
EcoR VCat.#FG-EcoRV

Size
4,000 units

Reaction conditions
Conc.
20 units/μl

1X FastGene® Buffer III, 37℃
1X FastGene® FastCut Buffer, 37℃

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

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

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

Unit definition
One unit is defined as the amount of enzyme required forcomplete digestion of 1 μg bacteriophage λ 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

Dilution buffer

Standard reaction condition

NIPPON Genetics
EUROPE GmbHFastGene® Diluent
A- Normal protocol

www.nippongenetics.eu
HeatInactivationwww.n-genetics.com

Substrate DNA

EcoR V can be inactivated at 65℃ for 20 min.

Methylation sensitivity

dam methylation: Not sensitive
dcm methylation: Not sensitive

Restriction Enzyme
CpG methylation: Conditionally sensitive
EcoR V
Prolonged incubation
A minimum
required to digest 1 μgConc.
substrateSizeCat.# amount of enzyme
DNAfor 16 hr; 0.5 U.
4,000 units
20 units/μl
FG-EcoRV®
Relative
Store atactivity-20℃ in FastGene Buffers

FastGene® Buffer I:0%100%FastGene® Buffer II:® Buffer III:FastGene100%® Buffer III
Suppliedwith:
10X
FastGene(FG-REB3)50%
FastGene® Buffer IV:® FastCut Buffer (FG-REBHF)
10X
FastGene
Buffer:100%
FastGene® FastCut 6X
DNA Loading Buffer

Note

Sterile water

No Recognition
further activity
is noted after 4 hr incubation at 37℃ andsite
thus overnight incubation is not effective. Cleavage ofmammalian DNA is blocked by overlapping CpG methylation. A
single EcoR V site is found in the tetracycline resistance gene ofpBR322.

Component

Volume1 μg

X μl1X
10X FastGene® Buffer III
Source: E.coli that carries the EcoR V gene
EcoR V from plasmid J62 pLG74 20 unit

5 μl

Sterile water

1 μl
up to 50 μl

Reaction conditions→ Incubate at 37℃ for 1 hr
1X FastGene® Buffer III, 37℃® FastCut Buffer, 37℃
1X FastGene- Fast protocol
Component

FastGene® FastCut Buffer

Final Conc.

Volume® restriction
SubstrateDNA
1 μgμlFastGene
enzyme can cut substrate
DNA in 5-15Xmin® FastCut®
with10X
FastGene
Buffer.
1X
5 μl
FastGene FastCut Buffer® Buffer IIIV1XEcoR
FastGene

20 unit

1 μl

50 SterilemM Tris-HCl
water (pH 7.9 at 25℃)
up to 50 μl
100 mM NaCl
→ Incubate at 37℃ for 15 min
10 mM MgCl2
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

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

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

Final Conc.

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