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Source: E. coli RY13

Restriction Enzyme
EcoRI

Reaction conditions
1X FastGene® Buffer EcoR I, 37℃
2X FastGene® FastCut Buffer, 37℃

FastGene® FastCut Buffer
Cat.#FG-EcoRI

Size
20,000 units

Conc.
20 units/μl

1X FastGene® Buffer EcoR I

Store at -20℃
Supplied with:

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

FastGene® 10XBuffer EcoR I
FastGene® 10X FastCut Buffer(FG-REBEcoRI)(FG-REBHF)

6X DNA Loading Buffer
Sterile water
Recognition site

100 mM Tris-HCl (pH 7.5 at 25℃)
50 mM NaCl
10 mM MgCl2
0.025% Triton X-100

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.

Dilution buffer

Quality control

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

FastGene® Diluent C- Unit definition assay- Overdigestion assay- Endonuclease assay- Extreme pure assay

NIPPON Genetics EUROPE GmbHHeat Inactivation

Standard reaction condition

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

- Normal protocol

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Methylation sensitivity

Component

dam methylation: Not sensitive
dcm methylation: Not sensitive

Substrate
DNARY13
Source:E. coliCpG methylation: Conditionally Sensitive

Restriction Enzyme
Prolonged incubation
A EcoRI
minimum amount of enzyme required to digest 1 μg substrate

Reaction
EcoR I conditions

DNA for 16 hr; 0.13 U.

FastGene® FastCut Buffer

Cat.#Size ®
Relative
activity in FastGene
Buffers

Conc.
FG-EcoRI
20,000 units
20 units/μl
FastGene® Buffer I:
50%
100%
FastGene® Buffer II:
Store at -20℃
FastGene® Buffer III:75%
® Buffer IV:
100%
FastGene® Buffer EcoR I
Suppliedwith:
10X
FastGene(FG-REBEcoRI)
Buffer:100%
FastGene® FastCut 10X® FastCut Buffer (FG-REBHF)
FastGene

6X DNA Loading Buffer
Sterile water
It can cleave non-specifically the sequences of AATT or
PuPuATPyPy (Pu=purine; Py=pyrimidine) (star activity). The
Recognition site
addition of 2 mM β-mercaptoethanol may inhibit this staractivity. Long-term storage can result in aggregation, leading toreduced activity. Both DNA and RNA in a DNA/RNA hybrid
duplex can be cleaved sequence specifically. Cleavage ofmammalian genomic DNA is impaired by overlapping CpG
methylation. Reaction condition of low salt, excess enzyme,
excess glycerol (>5%) or high pH (>8.0) can result in star activity.

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

Final Conc.

10X FastGene® Buffer EcoR I
1X FastGene® Buffer EcoR I, 37℃® FastCut Buffer, 37℃
Sterile water
2X FastGene

Volume1 μg

X μl1X5 μl

20 unit

1 μl
up to 50 μl→ Incubate at 37℃ for 1 hr

® restriction enzyme can cut substrate DNA in 5-15 min
FastGene- Fastprotocol
with FastGene® FastCut Buffer.
ComponentFinal Conc.
Volume® Buffer EcoR I
1X Substrate
FastGeneDNA

100 mM Tris-HCl ®(pH 7.5 at 25℃)
10X FastGene FastCut Buffer
50 mM NaCl
10 mM
EcoRMgCl
I 2
0.025% Triton X-100
Sterile water

1 μg

X μl

2X

10 μl

20 unit

1 μlup to 50 μl

Incubate at 37℃ for 15 min
Unit→definition
One unit is defined as the amount of enzyme required for※We digestion
recommend
units of enzyme
per37℃µg DNA
completeof 15-10
μg bacteriophage
λ atfor 1 andhr in10units forgenomic DNA in a 1 h digest.
50 μl20
reaction
mixtures.

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

Dilution buffer
FastGene® Diluent C