Thermo Scientific™ BspPI (AlwI) (2 U/μL)

Description

Conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

5'...G G A T C (N)₄▵...3'

3'...C C T A G (N)₅▵...5'

Conditions for 100% Activity:

• 1X Buffer Tango™: 33mM Tris-acetate (pH7.9 at 37°C), 10mM Mg-acetate, 66mM K-acetate and 0.1mg/mL BSA
• Incubate at 55°C

Storage Buffer:

• BspPI is supplied in:10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol

Ligation and Recleavage:

• After 10-fold overdigestion with BspPI, approximately 70% of the DNA fragments can be ligated in a reaction mixture containing 20-40U of T4 DNA Ligase/1μg of fragments and 10% PEG. More than 50% of these can be recut

Methylation Effects:

• Dam: completely overlaps — blocked
• Dcm: may overlap — no effect
• CpG: may overlap — no effect
• EcoKI: never overlaps — no effect
• EcoBI: may overlap — effect not determined

Note:

Incubation at 37°C results in 30% activity. Assayed using lambda DNA (dam) (#SD0021). BspPI is blocked by overlappingdam methylation. To avoiddam methylation, use adam-, dcm strain such as GM2163 (#M0099).