Store at -20℃. For Research Use Only!

dcm   CpG

Sau96  I

AspS9 I, Cfr13 I, PspP I

Supplied with : 10X Reaction Buffer 'C'

Concentration :            units/㎕

Lot # :                

Expiration date :                

Source : Staphylococcus  aureus  PS96 (ATCC 49831)

Compatible ends (5'…GNC)

AspS9 I, Asu I, BsiZ I, Cfr13 I, EcoO109 I, PspP I, Dra II

Isoschizomers : AspS9 I, Asu I, BsiZ I, Cfr13 I, PspP I

- neoschizomers : BmgT120 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 Sau96 I required to completely digest 1 ㎍ of unmethylated λ DNA in one hour at 37℃ in 50 ㎕ assay mixture.

Typical Reaction Conditions : Add 5 ㎕ of 10X Reaction Buffer 'C'[final conc.: 10 mM Tris-HCl(pH 7.9 @ 37℃), 10 mM MgCl₂, 50 mM NaCl, 1 mM DTT], 1~2 ㎕ of DNA(0.5~1.0 ㎍/㎕), 1 ㎕ of Sau96 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	10~50	75~100	50~75

Activity in a complete PCR Mix : 50~75%
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 Sau96 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
74	164	6	2	15	4	11

Quality control

Overdigestion: The same band pattern as a digestion in one unit of Sau96 I for one hour was showing after incubation of 1 ㎍ of λ DNA with         units of Sau96 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 Sau96 I.

Notes

- 1 µg pBR322 DNA is digested completely by ca. 3 units of Sau96 I on account of the larger number of cleavage sites per µg of

pBR322 DNA as compared to λ DNA.

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.