Store at -20℃. For Research Use Only!

CpG   ★

Sau3A  I

Bsp143 I, Dpn II, Kzo9 I, Mbo I, Nde II

Supplied with : 10X Reaction Buffer 'B'

Concentration :            units/㎕

Lot # :                

Expiration date :                

Source : Staphylococcus  aureus  3A (ATCC 49834)

Compatible ends (5'…GATC)

BamH I, BfuC I, Bcl I, Bgl II, Bsp143 I, BstMB I, BstX2 I, BstY I, Dpn II, Fba I, Ksp22 I, Kzo9 I, Mbo I, Mfl I, Nde II, Psu I, Xho II

Isoschizomers

: BfuC I, Bsp143 I, BssMI, BstMBI, DpnII, Kzo9 I, Mbo I, NdeII

- neoschizomer : BstKT 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 Sau3A 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 Sau3A I and 42~43 ㎕ of sterile water in 50 ㎕ reaction mixture. Incubate at 37℃.

Heat Inactivation : 65℃ for 20 min.

Activity in BeamsBio™ Buffer System

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

Activity in a complete PCR Mix : 75~100%
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 Sau3A I. And then incubate at 37℃ for one hour. Relative activity is determined by gel electrophoresis.

Storage Conditions : 10 mM Tris-HCl(pH 7.5 @ 25℃), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.15% Triton™ X-100, 500 ㎍/㎖ BSA, and 50% glycerol. Store at -20℃.

No. of Cleavage Sites

λ	Ad2	pUC18	φX174	pBR322	M13mp18	SV40
116	87	15	0	22	7	8

Quality control

Overdigestion: The same band pattern as a digestion in one unit of Sau3A I for one hour was showing after incubation of 1 ㎍ of λ DNA with         units of Sau3A 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 Sau3A I.

Notes

- Conditions of low ionic strength, high enzyme concentration, glycerol > 5%, or pH > 8.0 may result in star activity.

The relaxed sequence specificities are GAGC or CATC.

- Complete digestion of 1 μg pBR322 DNA requires ca. 5 U of Sau3A I because 1 μg pBR322 DNA has more cleavage sites

than 1 μg λ DNA.

- Isoschizomers of Sau3A I are Nde II and Mbo I. Mbo I and Nde II are sensitive to dam methylation.

References

Sussenbach, J. S., Steebergh, P. H., Rost, J. A., van Leeuwen, W. J. and van Embden, J. D. A. Nucleic Acids Res. 5: 1153-1163 (1978)

Beams Biotechnology Co., Ltd.