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Hind III (10KU/vl Cat.#1025), "B" buffer
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Store at -20¡É. For Research Use Only!

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Hind  III

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

#1025

10,000 units

#1025L

50,000 units

Supplied with : 10X Reaction Buffer 'B'


Concentration :            units/§¡


Lot # :                


Expiration date :                


Source : Haemophilus influenzae Rd (exo- mutant)

(ATCC 51907)


Compatible ends (5'¡¦AGCT) & Isoschizomers

: Bbr I, BspKT8 I, BstF I, EcoV ¥², Hsu I, LlaC I, Ssb I


Methylation Sensitivity

: Not sensitive to dam, dcm and CpG methylation


Unit definition : One Unit is defined as the amount of Hind ¥² 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 Hind ¥² 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

75~100

75~100

10~50



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 Hind ¥². 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 @ 25¡É), 400 mM KCl, 0.1 mM EDTA, 1 mM DTT, 100 §¶/§¢ BSA, and 50% glycerol. Store at -20¡É.



No. of Cleavage Sites

¥ë

Ad2

pUC18

¥õX174

pBR322

M13mp18

SV40

6

12

1

0

1

1

6



Quality control

Overdigestion: The same band pattern as a digestion in one unit of Hind ¥² for one hour was showing after incubation of 1 §¶ of ¥ë DNA with         units of Hind ¥² 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 Hind ¥².


Notes

- Star activity may be resulted in the presence of Mn2+.

- The activity of Hind ¥² may be enhanced with the addition of BSA at a concentration of 100 §¶/§¢ without any negative effect.


References

Hsu, M., and Berg, P. Biochemistry. Biochemistry 17: 131-138, (1978)/ Old, R., Murray, K., Roizes, G. J. Mol. Biol. 92: 331-339 (1975)


Beams Biotechnology Co., Ltd.


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