Thermo Scientific™ PagI (BspHI)

The PagI (BspHI) restriction enzyme recognizes T^CATGA sites and cuts best at 37°C in O buffer (Isoschizomers: BspHI, CciI, RcaI).

Brand:  Thermo Scientific™ ER1281

Product Code. 10809870

Description

5'...T▵C A T G A...3'

3'...A G T A C▵T...5'

Conditions for 100% Activity:

• 1X Buffer O: 50mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl₂, 100mM NaCl and 0.1mg/mL BSA
• Incubate at 37°C

Storage Buffer:

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

Ligation and Recleavage:

• After 50-fold overdigestion with PagI, more than 95% of the DNA fragments can be ligated and recut

Methylation Effects:

• Dam: may overlap —cleavage impaired
• Dcm: never overlaps —no effect
• CpG: never overlaps —no effect
• EcoKI: never overlaps —no effect
• EcoBI: may overlap —cleavage impaired

Digestion of Agarose-embedded DNA:

• Minimum 5units of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours

Compatible Ends:

• AflIII, PscI, DsaI, Eco130I, NcoI

Note:

Low salt, high glycerol (>5%) concentrations, pH >8.0 or a large excess of enzyme may result in star activity. PagI cleavage is impaired by overlapping dam methylation. To avoid dam methylation, use a dam-, dcm strain such as GM2163 (#M0099).

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Specifications

• Product Name: PagI (BspHI)
• Concentration: 10U/µL
• Units: 400U