Store at -20℃. For Research Use Only!

CpG   ★

Ava  I

Ama87 I, BsoB I, Eco88 I

Supplied with : 10X Reaction Buffer 'B'

Concentration :            units/㎕

Lot # :                

Expiration date :                

Source : Anabaena variabilis ATCC 27893

Compatible ends (5'…YCGR)

Ama87 I, Bco I, BsoB I, BsiHKC I, Eco88 I, Nsp III

Isoschizomers

: Ama87 I, BmeT110 I(neoschizomer), BsiHKC I, BsoB I, Eco88 I

Methylation Sensitivity

: Not sensitive to dam and dcm methylation

: Blocked by CpG methylation

Unit definition : One Unit is defined as the amount of Ava I required to completely digest 1 ㎍ of λ DNA in one hour at 37℃ in 50 ㎕ assay mixture.

Typical Reaction Conditions : Add 5 ㎕ of 10X Reaction Buffer 'B'[final conc.: 6 mM Tris-HCl(pH 7.5 @ 37℃), 6 mM MgCl₂, 50 mM NaCl, 1 mM DTT], 1~2 ㎕ of DNA(0.5~1.0 ㎍/㎕), 1 ㎕ of Ava I and 42~43 ㎕ of sterile water in 50 ㎕ reaction mixture. Incubate at 37℃.

Heat Inactivation : 80℃ for 20 min.

Activity in BeamsBio™ Buffer System

Rxn Buffer	A	B	C	D
Activity(%)	10~50	75~100	50~75	10~50

Activity in a complete PCR Mix : 10~50%
Conditions: [10 mM Tris-HCl(pH 8.3 @ 25℃), 50 mM KCl, 1.5 mM MgCl₂, 2 pmol of primers, 200 μM 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 I. And then incubate at 37℃ for one hour. Relative activity is determined by gel electrophoresis.

Storage Conditions : 10 mM Tris-HCl(pH 7.4 @ 25℃), 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, 200 ㎍/㎖ BSA, and 50% glycerol. Store at -20℃.

No. of Cleavage Sites

λ	Ad2	pUC18	φX174	pBR322	M13mp18	SV40
8	40	1	1	1	2	0

Quality control

Overdigestion: The same band pattern as a digestion in one unit of Ava I for one hour was showing after incubation of 1 ㎍ of λ DNA with         units of Ava I for 16 hr in 50 ㎕ of reaction mixture at 37℃ as determined by agarose gel electrophoresis.

Ligation-Recut: After a 10-fold digestion for one hour,        % of λ DNA fragments were ligated with T4 DNA ligase at 16℃ and        % of the ligated fragments could be recut with Ava I.

Notes

- Conditions of glycerol concentration > 5% (v/v) or the presence of DMSO may result in star activity.

- Ava I requires at least 2 base pairs beyond the end of its recognition sequence to cleave efficiently.

- Activity is enhanced 2.5-fold by incubation at 45°C.

References

Murray, K., Hughes, S.G., Brown, J.S., Bruce, S.A., Biochem. J., 159: 317-322 (1976)/ Hughes, S.G., Murray, K., Biochem. J., 185: 65-75 (1980)/ Choi, W.S., Kang, S.C., Seo, J.S., Yoo, O.J., Korean J. Microbiol., 24: 86-90 (1986)

Beams Biotechnology Co., Ltd.