Thermo Scientific™ BspPI (AlwI)

Cut at GGATC(4/5)^ sites with BspPI (AlwI) restriction enzyme, which performs best at 55°C in Tango buffer (Isoschizomers: AclWI, AlwI).

Brand:  Thermo Scientific™ ER1321

Product Code. 10384630

Description

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).

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Specifications

• Product Name: BspPI (AlwI)
• Concentration: 1 to 3U/µL
• Incubator Temperature: 55°C
• Quantity: 100U