Store at -20℃. For Research Use Only!

Bgl  II

Supplied with : 10X Reaction Buffer 'D'

Concentration :            units/㎕

Lot # :                

Expiration date :                

Source : Bacillus  globigii  ATCC 49760

Compatible ends (5'…GATC)

BamH I, Bcl I, BfuC I, Bsp143 I, BstMB I, BstX2 I, BstY I, Dpn II, Fba I, Ksp22 I, Kzo9 I, Mbo I, Mfl I, Nde II, Psu I, Sau3A I, Xho II

Isoschizomers

: Ncr I

Methylation Sensitivity

: Not sensitive to dcm, dam and CpG methylation

Unit definition : One Unit is defined as the amount of Bgl II 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 'D'[final conc.: 6 mM Tris-HCl(pH 7.9 @ 37℃), 6 mM MgCl₂, 150 mM NaCl, 1 mM DTT], 1~2 ㎕ of DNA(0.5~1.0 ㎍/㎕), 1 ㎕ of Bgl II and 42~43 ㎕ of sterile water in 50 ㎕ reaction mixture. Incubate at 37℃.

Heat Inactivation : No

Activity in BeamsBio™ Buffer System

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

Activity in a complete PCR Mix : less than 10%
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 Bgl 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℃), 50 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 500 ㎍/㎖ BSA, and 50% glycerol. Store at -20℃.

No. of Cleavage Sites

λ	Ad2	pUC18	φX174	pBR322	M13mp18	SV40
6	11	0	0	0	1	0

Quality control

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

Notes

- Bgl II is not inactivated by heating and therefore should be extracted with phenol/chloroform and once with chloroform,

and then precipitate the DNA with ethanol.

References

Pirrotta, V. Nucleic Acids Res. 3: 1747-1760 (1976)/ Duncan, C.H., Wilson, G.A. and Young, F.E. J. Bacteriol. 134: 338 (1978)/ Bickle, T.A., Pirrotta, V., Imber, R. Methods Enzymol. 65: 132-138 (1980)

Beams Biotechnology Co., Ltd.