Store at -20℃. For Research Use Only!

CpG   ★

EcoR  I

Rsr I

Supplied with : 10X Reaction Buffer 'EcoR I'

Concentration :            units/㎕

Lot # :                

Expiration date :                

Source : Escherichia  coli  RY13

Compatible ends (5'…AATT)

: Acs I, Apo I, Mfe I, Mun I, Sse9 I, Tas I, Tsp509 I, Xap I

Isoschizomers : Rsr I

Methylation Sensitivity

: Not sensitive to dam and dcm methylation

: Affected by overlapping CpG methylation

Unit definition : One Unit is defined as the amount of EcoR I required to completely digest 1 ㎍ of λ DNA in one hour at 37℃ in 50 ㎕ assay mixture.

Typical Reaction Conditions : Add 5 ㎕ of 10X Reaction Buffer 'EcoR I'[final conc.: 90 mM Tris-HCl (pH 7.5 @ 37℃), 10 mM MgCl₂, 50 mM NaCl], 1~2 ㎕ of DNA(0.5~1.0 ㎍/㎕), 1 ㎕ of EcoR I and 42~43 ㎕ of sterile water in 50 ㎕ reaction mixture. Incubate at 37℃.

Heat Inactivation : 65℃ for 20 min.

Activity in BeamsBio™ Buffer System

Rxn Buffer	A	B	C	D	'EcoR I'
Activity(%)	10~50	50~75	50~75	50~75	75~100

Activity in a complete PCR Mix : 10~50%
Conditions: [10 mM Tris-HCl(pH 8.3 @ 25 ℃), 50 mM KCl, 1.5 mM MgCl₂, 2 pmol of primers, 200 μM dNTP Mix, 2.5 U Taq DNA polymerase and 1 ㎍ of DNA(template or substrate) in 50 ㎕ of reaction volume] After 30 amplication cycles, add 5 units of EcoR I. And then incubate at 37℃ for one hour. Relative activity is determined by gel electrophoresis.

Storage Conditions : 10 mM Tris-HCl(pH 7.5), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.15% Triton™ X-100, 100 ㎍/㎖ BSA, and 50% glycerol. Store at -20℃.

No. of Cleavage Sites

λ	Ad2	pUC18	φX174	pBR322	M13mp18	SV40
5	5	1	0	1	1	1

Quality control

Overdigestion: The same band pattern as a digestion in one unit of EcoR I for one hour was showing after incubation of 1 ㎍ of λ DNA with         units of EcoR I for 16 hr in 50 ㎕ of reaction mixture at 37℃ as determined by agarose gel electrophoresis.

Ligation-Recut: After a 10-fold digestion for one hour,        % of λ DNA fragments were ligated with T4 DNA ligase at 16℃ and        % of the ligated fragments could be recut with EcoR I.

Notes

- Conditions of low ionic strength, high enzyme concentration, glycerol concentration > 5%, or pH > 8.0 may result in star activity.

References

Hedgpeth, J., Goodman, H.M., Boyer, H.W. Proc. Natl. Acad. Sci. U. S. A. 69: 3448-3452 (1972)/ Albertsen, H.M., Le Paslier, D., Abderrahim, H., Dausset, J., Cann, H., Cohen, D. Nucleiv Acids Res. 17: 808 (1989)

Beams Biotechnology Co., Ltd.