Product Class: Restriction Endonuclease

Nb.BtsI

Description

Nb.BtsI is a nicking endonuclease that cleaves only one strand of DNA on a double- stranded DNA substrate.

Product Source

An E. coli strain that expresses only the large subunit of the BtsI restriction gene from Bacillus thermoglucosidasius (X.Pan).

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
CutSmart® Buffer-2010X

Properties and Usage

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.

Reaction Conditions

1X CutSmart® Buffer
Incubate at 37°C

1X CutSmart® Buffer:
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 μg/ml BSA
pH 7.9 @ 25°C

Activity in NEBuffers

NEBuffer 1.1: 75%
NEBuffer 2.1: 100%
NEBuffer 3.1: 75%
CutSmart™ Buffer: 100%

Diluent Compatibility

Storage Temperature

-20°C

Storage Conditions

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

Heat Inactivation

80°C for 20 min

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive

Heat Inactivated

Yes

Quality Control

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
  • Exonuclease Activity (Radioactivity Release):
    The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
  • Non-Specific DNase Activity (16 hour):
    The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.

Supporting Documents

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Notes

  1. 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. 
  2. Nb.BtsI is 100% active at 55°C. 
  3. To run on an electrophoresis gel, add loading dye containing a final concentration of 0.4% SDS.

References

  1. Song, Q. et al. (2010). Anal. Chem. [Epub ahead of print].
  2. Zhang, P. et al. (2010). Protein Expr. Purif. 69, 226-234. [Epub 2009 Sep 9].
  1. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
  2. How can I access the old NEBuffer Activity Chart?
  3. How can I access the old Double Digest Finder?
  1. Optimizing Restriction Endonuclease Reactions
  2. Double Digest Protocol with Standard Restriction Enzymes

Selection Tools

Usage Guidelines & Tips

Troubleshooting Guides

Interactive Tools

Also has 100% activity in NEBuffer 2
Add 0.4% SDS before running agarose gel to dissociate DNA-enzyme complex