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BsmBI
Cloned At NEBRecombinant SourceNEBuffer 355Heat Inactivated
Catalog # Size Concentration Price Qty  
R0580L 1,000 units 10,000 units/ml $264.00
R0580S 200 units 10,000 units/ml $66.00
Prices are in US dollars and valid only for US orders.
Download:MSDS PDF


Recognition Site:

CGTCTC

isoschizomers | compatible ends | single letter code

Source:
An E.coli strain that carries the BsmBI gene from Bacillus stearothermophilus B61 (Z. Chen).

Reagents Supplied:
NEBuffer 3


Enzyme Properties


Activity in NEBuffers:
NEBuffer 1:75%
NEBuffer 2:100%
NEBuffer 3:100%
NEBuffer 4:100%

When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion.

Methylation Sensitivity:
dam methylation: Not sensitive
dcm methylation: Not sensitive
CpG methylation: Blocked
More information about: Methylation Sensitivity

Activity at 37°C:
20%

Heat Inactivation:
80°C for 20 minutes

Survival in a Reaction:
(+) Intermediate activity. Suitable for extended digestion, but < 8 hours.
More information about: Extended Digests with Restriction Enzymes


Reaction & Storage Conditions


Reaction Conditions:
1X NEBuffer 3
Incubate at 55°C.

1X NEBuffer 3:
50 mM Tris-HCl
100 mM NaCl
10 mM MgCl2
1 mM Dithiothreitol
pH 7.9 @ 25°C

Unit Definition:
One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 55°C in a total reaction volume of 50 µl.

Concentration:
10,000 units/ml

Unit Assay Substrate:
λ DNA

Storage Conditions:
10 mM Tris-HCl
200 mM NaCl
1 mM Dithiothreitol
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Storage Temperature:
-20°C

Diluent Compatibility:
Diluent B


Notes


General notes:
  1. BsmBI is an isoschizomer of Esp3I.

FAQs


  1. Why is there a BsmBI site in LITMUS 38 but not LITMUS 39?

Quality Control for Current Lot


Quality control values for a specific lot can be found on the datacard which accompanies each product.

Ligation and Re-cutting:
After a 10-fold overdigestion with BsmBI, > 95% of the DNA fragments can be ligated with T4 DNA Ligase (at a 5' termini concentration of 1-2 μM) at 16ºC. Of these ligated fragments, > 95% can be recut with BsmBI.


16-Hour Incubation:
A 50 μl reaction containing 1 μg of DNA and 10 units of BsmBI incubated for 16 hours at 55ºC resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.

Exonuclease Activity:
Incubation of a 50 μl reaction containing 60 units of BsmBI with 1 μg of a mixture of single and double-stranded [3H] E. coli DNA (205 cpm/μg) for 4 hours at 55ºC released < 0.1% of the total radioactivity.

Endonuclease Activity:
Incubation of a 50 μl reaction containing 10 units of BsmBI with 1 μg of ΦX174 RF I DNA for 4 hours at 55ºC resulted in < 10% conversion to RFII as determined by agarose gel electrophoresis.


Reagents Sold Separately


NEBuffer 3


Legal


Patents:
New England Biolabs, Inc.: U.S. Patent No. 6,764,843

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