Product Source

Bacillus stearothermophilus

Reagents Supplied

The following reagents are supplied with this product: 

Unit Definition

One unit is defined as the amount of enzyme required to convert 1 μg of supercoiled pUC19 DNA to open circular form in 1 hour at 37°C in a total reaction volume of 50 μl.

Reaction Conditions

1X NEBuffer 3.1
Incubate at 37°C

1X NEBuffer 3.1:
100 mM NaCl
50 mM Tris-HCl
10 mM MgCl₂
100 μg/ml BSA
pH 7.9 @ 25°C

Activity in NEBuffers

NEBuffer 1.1: 10%
NEBuffer 2.1: 100%
NEBuffer 3.1: 100%
CutSmart® Buffer: 25%

Diluent Compatibility

• Diluent B

Storage Temperature

-20°C

Storage Conditions

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

Heat Inactivation

No

Methylation Sensitivity

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

Long Term Storage

• 
  Shelf Life 2 years

Companion Products

• Monarch® Plasmid Miniprep Kit 
• Monarch® DNA Gel Extraction Kit
• Monarch® PCR & DNA Cleanup Kit (5 μg)

Materials Sold Separately

• NEBuffer 3.1

1.  Incubation at 50°C results in 3 fold higher activity.

2.  For enzymes that cannot be heat-inactivated, we recommend using a column for cleanup (such as the Monarch® PCR & DNA Cleanup Kit), or running the reaction on an agarose gel and then extracting the DNA (we recommend Monarch Gel Extraction Kit), or performing a phenol/chloroform extraction.