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Ava II (1KU/vl Cat.#1006), "C" buffer
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  Store at -20¡É. For Research Use Only!

dcm   CpG   ¡Ú

Ava  II

Bme18 I, Eco47 I, Sin I

5' ¡¦ G G (A/T) C C ¡¦ 3'
3' ¡¦ C C (T/A) G G ¡¦ 5'

#1006

1,000 units

#1006L

5,000 units

Supplied with : 10X Reaction Buffer 'C'


Concentration :            units/§¡


Lot # :                


Expiration date :                


Source : Anabaena variabilis ATCC 27893


Compatible ends (5'¡¦GWC)

Bme18 I, Cpo I, Csp I, Eco47 I, PpuM I, Psp5 II, PspPP I, Rsr II, Rsr2 I, SanD I, Sin, VpaK11B I


Isoschizomers

: Bme18 I, Eco47 I, Sin I, VpaK11B I


Methylation Sensitivity

: Not sensitive to dam methylation

: Blocked by overlapping dcm methylation

: Blocked by overlapping CpG methylation


Unit definition : One Unit is defined as the amount of Ava 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 'C'[final conc.: 10 mM Tris-HCl(pH 7.9 @ 37¡É), 10 mM MgCl2, 50 mM NaCl, 1 mM DTT], 1~2 §¡ of DNA(0.5~1.0 §¶/§¡), 1 §¡ of Ava II 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(%)

50~75

50~75

75~100

10~50



Activity in a complete PCR Mix : less than 10%
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 Ava II. And then incubate at 37¡É for one hour. Relative activity is determined by gel electrophoresis.


Storage Conditions : 10 mM Tris-HCl(pH 7.4 @ 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

35

73

2

1

8

1

6


Quality control

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


Notes

- High glycerol concentration may result in star activity.


References

Murray, K., Hughes, S.G., Brown, J.S., Bruce, S.A., Biochem. J., 159: 317-322 (1976)/ Sutcliffe, J. G., Church, G. M., Nucleic Acids Res., 5: 2313-2319 (1978)/ Fuchs, C., Rosenvld, E. C., Honigman, A., Szybalsky, W., Gene, 4: 1-23 (1978)


Beams Biotechnology Co., Ltd.


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