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Dde I (200U/vl Cat.#1016), "D" buffer
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  Store at -20¡É. For Research Use Only!

Dde  I

BstDE I, HpyF3 I

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

#1016

200 units

#1016L

1,000 units

Supplied with : 10X Reaction Buffer 'D'


Concentration :            units/§¡


Lot # :                


Expiration date :                


Source : Desulfovibrio desulfuricans  (NCIB 83210)


Compatible ends (5'¡¦TNA)

Axy I, Blp I, Bpu10 I, Bpu1102 I, Bse21 I, Bsp1720 I, Bsu36 I, BstDE I, Cel II, Eco81 I, HpyF3 I


Isoschizomers : BstDE I, HpyF3 I


Methylation Sensitivity

: Not sensitive to dam, dcm and CpG methylation


Unit definition : One Unit is defined as the amount of Dde 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 'D'[final conc.: 6 mM Tris-HCl(pH 7.9 @ 37¡É), 6 mM MgCl2, 150 mM NaCl, 1 mM DTT], 1~2 §¡ of DNA(0.5~1.0 §¶/§¡), 1 §¡ of Dde 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(%)

10~50

10~50

50~75

75~100



Activity in a complete PCR Mix : 50~75%
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 Dde 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), 50 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

104

97

6

14

8

29

20


Quality control

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


Notes

- Single-stranded DNA cleaves slowly.


References

Makula, R.A., Meagher, R.B. Nucleic Acids Res., 8: 3125-3131 (1980)


Beams Biotechnology Co., Ltd.


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