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Hinf I (3KU/vl Cat.#1026), "B" buffer
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  Store at -20¡É. For Research Use Only!

CpG   ¡Ú

Hin I

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

#1026

3,000 units

#1026L

15,000 units

Supplied with : 10X Reaction Buffer 'B'


Concentration :            units/§¡


Lot # :                


Expiration date :                


Source : Haemophilus influenzae Rf (ATCC 17947)


Compatible ends (5'¡¦ANT) & Isoschizomers

: CviB I, FnuA I, Hha II, SscL1 I


Methylation Sensitivity

: Not sensitive to dam and dcm methylation

: Blocked by overlapping CpG methylation


Unit definition : One Unit is defined as the amount of Hinf 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 Hinf I and 42~43 §¡ of sterile water in 50 §¡ reaction mixture. Incubate at 37¡É.


Heat Inactivation : 80¡É for 20 min.


Activity in BeamsBio¢â Buffer System

Rxn Buffer

A

B

C

D

Activity(%)

50~75

75~100

75~100

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 Hinf 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 @ 25¡É), 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

148

72

6

21

10

27

10


Quality control

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


Notes

- Hinf I requires optimal reaction conditions in order to avoid star activity.


References

Godson, G.N., Roberts, R.J. Virology 73: 561-567 (1976)/ Blakesley, R.W., et al. J. Biol. Chem. 252: 7300-7306 (1977)


Beams Biotechnology Co., Ltd.

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