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Restriction Endonucleases >
Nicking Endonucleases >
Nb.BtsI |
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Prices are in US dollars and valid only for US orders.
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Recognition Site:
isoschizomers | single letter code
Description:
Nb.BtsI is a nicking endonuclease that cleaves only one strand of DNA on a double- stranded DNA substrate.
Source:
An E. coli strain that expresses only the large subunit of the BtsI restriction gene from Bacillus thermoglucosidasius (X.Pan).
Reagents Supplied:
NEBuffer 4 (10X)
BSA (100X)
Enzyme Properties
Activity in NEBuffers:
| NEBuffer 1: | | 75% | | NEBuffer 2: | | 100% | | NEBuffer 3: | | 75% | | 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.
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 4
Supplemented with 100 μg/ml Bovine Serum Albumin
Incubate at
37°C.
1X NEBuffer 4:
20 mM Tris-acetate
50 mM potassium acetate
10 mM Magnesium Acetate
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 ΦX174 RF I DNA to open circular form in 1 hour at 37°C in a total reaction volume of 50 µl.
Concentration:
10,000 units/ml
Unit Assay Substrate:
ΦX174 DNA
Storage Conditions:
10 mM Tris-HCl
50 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 A
Notes
General notes:- Nb.BtsI is very efficient. Under most conditions a one to two hour incubation with 1 µl of enzyme and 1 µg of DNA is recommended.
Nb.BtsI is 100% active at 55°C.
To run on an electrophoresis gel, add loading dye containing a final concentration of 0.4% SDS.
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.BtsI 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.BtsI 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 4
BSA
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