Izoschizomers

Bst2B I

Enzyme name

Bst2B I

Prototype

BsiI

Known analogs

BauI, BsiI, BssSI

SKU

SE-E043

Turbo version

Not available

High-concentration version

Not available

Recognition site

5'… C^▼TCGTG …3'

3'… GAGCA_▲C …5'

Source

Bacillus stearothermophilus 2B

Optimal buffer

SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)

Optimal temperature

60 °C

Inactivation temperature

80 °C

Buffer activity

B	G	O	W	Y	ROSE
75	25	10	25	100	100

Unit definition

One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 60°C in a total reaction volume of 50 μl.

Assayed on

Lambda DNA

Storage conditions

10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol. Store at -20°C.

Ligation

After 10-fold overdigestion with enzyme 95% of the DNA fragments can be ligated and recut.

Nonspecific hydrolysis

No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 5 u.a. of enzyme for 16 hours at 60°C.

Methylation sensitivity

not tested

Reagents supplied with enzyme

10 X SE-buffer Y, BSA

Notes

To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml. Do not use BSA for long incubation.

References

Unpublished data Abdurashitov, M.A., Belichenko, O.A., Degtyarev, S.K Unpublished observations (1995). 1995
V.S. Dedkov, N.I. Rechkunova, S.Kh. Degtyarev, P.A. Zhilkin, A.A. Kolykhalov, V.A. Verkhozina Bacillus sphaericus - a strain producer of restriction endonuclease BsiI. // Soviet Union patent SU 1784642 (1992). (In Russian) 1992
S.Kh. Degtyarev, A.A. Kolykhalov, N.I. Rechkunova, V.S. Dedkov, P.A. Zhilkin BsiI - novel unusual restriction endonuclease. // Molecular Biology(Moscow), Vol.24, No. 1, 244-247 (1990). (In Russian) 1990