Store at -20℃. For Research Use Only!

CpG

Cla  I

Bsa29 I, BspD I, BspX I, Bsu15 I

Supplied with : 10X Reaction Buffer 'C'

Concentration :            units/㎕

Lot # :                

Expiration date :                

Source : Caryophanon  latum  L  (ATCC 49862)

Compatible ends (5'…CG)

Aci I, Acy I, Acl I, Ban III, BmeT110 I, Bpu14 I, Bsa29 I, BsaH I, BseC I, BshV I, Bsp119 I, BspAC I, BspD I, BspT104 I, BspX I, BstAC I, BstB I, Bsu15 I, BsuTU I, Csp45 I, Hap II, Hin1 I, Hin6 I, HinP1 I, Hpa II, HpyCH4 Ⅳ, HspA I, Hsp92 I, Mae II, Mly113 I, Msp I, Nar I, Sfu I, Ssi I, Taq I

Isoschizomers

: Ban Ⅲ, Bsa29 I, BseC I, BshV I,

BspD I, BspX I, Bsu15 I, BsuTU I

Methylation Sensitivity

: Blocked by overlapping dam methylation

: Not sensitive to dcm methylation

: Blocked by CpG methylation

Unit definition : One Unit is defined as the amount of Cla I required to completely digest 1 ㎍ of λ(dam^-) 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 Cla I and 42~43 ㎕ of sterile water in 50 ㎕ reaction mixture. Incubate at 37℃.

Heat Inactivation : 65℃ for 20 min.

Storage Conditions : 10 mM Tris-HCl(pH 7.4 @ 25℃), 50 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 500 ㎍/㎖ BSA, and 50% glycerol. Store at -20℃.

Activity in BeamsBio™ Buffer System

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

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 Ava II. And then incubate at 37℃ for one hour. Relative activity is determined by gel electrophoresis.

No. of Cleavage Sites

λ	Ad2	pUC18	φX174	pBR322	M13mp18	SV40
15	2	0	0	1	2	0

Quality control

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

Notes

- Cla I requires 5 units to cleave 1 ㎍ of pBR322 DNA.

References

Mayer, H., Grosschedl, R., Schutte, H., Hobom, G., Nucleic Acids Res., 9, 4833-4845, (1981)

Beams Biotechnology Co., Ltd.