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Alu I (400U/vl Cat.#1003), "B" buffer
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  Store at -20¡É. For Research Use Only!

Alu  I

AluB I

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

#1003

400 units

#1003L

2,000 units

Supplied with : 10X Reaction Buffer 'B'


Concentration :            units/§¡


Lot # :                


Expiration date :                


Source : Arthrobacter  luteus  ATCC 21606


Compatible ends : with any blunt end


Isoschizomers : AluB I


Methylation Sensitivity

: Not sensitive to dam, dcm and CpG methylation


Unit definition : One Unit is defined as the amount of Alu 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 'B'[final conc.: 6 mM Tris-HCl(pH 7.5 @ 37¡É), 6 mM MgCl2, 50 mM NaCl, 1 mM DTT], 1~2 §¡ of DNA(0.5~1.0 §¶/§¡), 1 §¡ of Alu 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

Activity(%)

75~100

75~100

75~100

10~50


Activity in a complete PCR Mix : 75~100%
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 Alu I. And then incubate at 37¡É for one hour. Relative activity is determined by gel electrophoresis.


Storage Conditions : 10 mM Tris-HCl(pH 7.4 @ 25¡É), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 200 §¶/§¢ BSA, and 50% glycerol. Store at -20¡É.


No. of Cleavage Sites

¥ë

Ad2

pUC18

¥õX174

pBR322

M13mp18

SV40

143

158

16

24

17

27

34



Quality control

Overdigestion: The same band pattern as a digestion in one unit of Alu I for one hour was showing after incubation of 1 §¶ of ¥ë DNA with         units of Alu 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 Alu I.


Notes

- The activity of Alu I is reduced in buffers containing salt concentrations above 100mM, so Alu I is 10~50% active

in BeamsBio¢â Buffer 'D'.


References

Roberts, R. J., Myers, P. A. Morrison, A., Murray, K., J. Mol. Biol., 102, 157 (1976)/ Kim, J.H., Boo, Y.B., Lee, K.Y., Korean J. Biochem., 17: 149-154 (1985)/ Butkus, V. et al., Biochim. Biophys. Acta, 909: 201-207 (1987)


Beams Biotechnology Co., Ltd.


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