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EcoR I (10KU/vl Cat.#1018), Àü¿ë buffer
»óÇ°°¡ 60,500¿ø(ºÎ°¡¼¼Æ÷ÇÔ)
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  Store at -20¡É. For Research Use Only!

CpG   ¡Ú

Eco I

Rsr I

5' ¡¦ G A A T T C ¡¦ 3'
3' ¡¦ C T T A A G ¡¦ 5'

#1018

10,000 units

#1018L

50,000 units

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 ¥ë DNA in one hour at 37¡É in 50 §¡ assay mixture.


Typical Reaction Conditions : Add 5 §¡ of 10X Reaction Buffer 'EcoI'[final conc.: 90 mM Tris-HCl (pH 7.5 @ 37¡É), 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¡É.


Heat Inactivation : 65¡É for 20 min.


Activity in BeamsBio¢â 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 ¡É), 50 mM KCl, 1.5 mM MgCl2, 2 pmol of primers, 200 ¥ìM 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¡É 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¢â X-100, 100 §¶/§¢ BSA, and 50% glycerol. Store at -20¡É.


No. of Cleavage Sites

¥ë

Ad2

pUC18

¥õX174

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 ¥ë DNA with         units of EcoR I for 16 hr in 50 §¡ of reaction mixture at 37¡É as determined by agarose gel electrophoresis.

Ligation-Recut: After a 10-fold digestion for one hour,        % of ¥ë DNA fragments were ligated with T4 DNA ligase at 16¡É 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.


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