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Pvu II (2KU/vl Cat.#1044), "B" buffer
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  Store at -20¡É. For Research Use Only!

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Pvu  II

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

#1044

2,000 units

#1044L

10,000 units

Supplied with : 10X Reaction Buffer 'B'


Concentration :            units/§¡


Lot # :                


Expiration date :                


Source : Proteus vulgaris  (ATCC 13315)


Compatible ends : with any blunt end


Isoschizomers : Not known.


Methylation Sensitivity

: Not sensitive to dam, dcm and CpG methylation


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


Heat Inactivation : No


Activity in BeamsBio¢â Buffer System

Rxn Buffer

A

B

C

D

Activity(%)

10~50

75~100

50~75

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 Pvu II. 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

15

24

2

0

1

3

3


Quality control

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


Notes

- Pvu II is not inactivated by heating and therefore should be extracted with phenol/chloroform and once with chloroform,

and then precipitate the DNA with ethanol.

- Pvu II exhibits star activity when used under conditions of low ionic strength, high enzyme concentration,

glycerol concentration > 5%, or pH > 8.0.


References

Gingeras, T.R., Greenough, L., Schildkraut, I., Roberts, R.J. Nucleic Acids Res. 9: 4525-4536 (1981)


Beams Biotechnology Co., Ltd.


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