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Restriction Endonucleases >
Nicking Endonucleases >
Nb.BsmI |
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Recognition Site:
isoschizomers | single letter code
Description:
Nb.BsmI is a nicking endonuclease that cleaves only one strand of DNA on a double-stranded DNA substrate.
Source:
An E. coli strain that carries the cloned BsmI gene from Bacillus stearothermophilus NUB 36 (N. Welker)
Reagents Supplied:
NEBuffer 3
Enzyme Properties
Activity in NEBuffers:
| NEBuffer 1: | | 25% | | NEBuffer 2: | | 100% | | NEBuffer 3: | | 100% | | NEBuffer 4: | | 25% |
When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Activity at 37°C:
25%
Heat Inactivation:
80°C for 20 minutes
Survival in a Reaction:
Minimum units to digest 1 µg of substrate DNA in 16 hours: 0.25 unit(s)
Reaction & Storage Conditions
Reaction Conditions:
1X NEBuffer 3
Incubate at
65°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 convert 1 µg of supercoiled plasmid pBR322 DNA to open circular form in 1 hour at 65°C in a total reaction volume of 50 µl.
Concentration:
10,000 units/ml
Unit Assay Substrate:
pBR322 DNA
Storage Conditions:
10 mM Tris-HCl
100 mM NaCl
1 mM Dithiothreitol
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C
Storage Temperature:
-20°C
Diluent Compatibility:
Diluent A
Notes
General notes:- Use of Nb.BsmI in NEBuffer 1 and NEBuffer 4 may result in cleavage of both DNA strands. Incubation times of > 4 hours are not recommended.
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
16-Hour Incubation:
A 50 μl reaction containing 1 μg of DNA
and 10 units of Nb.BsmI incubated for 16 hours at 37ºC
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 100 units of Nb.BsmI with 1 μg of a mixture of single and double-stranded
[3H] E. coli DNA (205 cpm/μg) for 4 hours at 37ºC
released < 0.1% of the total radioactivity.
Reagents Sold Separately
NEBuffer 3
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