Thermo Scientific FspBI (BfaI)
The FspBI (BfaI) restriction enzyme recognizes C^TAG sites and cuts best at 37°C in Tango buffer (Isoschizomers: BfaI, MaeI, XspI).
Manufacturer: Thermo Scientific ER1762
Lambda DNA digested with FspBI (BfaI), 0.7% agarose, 13 cleavage sites
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.
More than 80% of the digested DNA fragments can be ligated and more than 95% can be recut in a reaction mixture containing 20 to 40U of T4 DNA ligase/1µg of fragments and 10% PEG at a 5ft.-termini concentration of 0.13µM after a 50-fold overdigestion(3U/µg DNA x 17 hours) with BfuI FspBI (BfaI).
5'...C▵T A G...3'
3'...G A T▵C...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 37°C
- FspBI 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
- Dam: may overlap no effect
- Dcm: never overlaps no effect
- CpG: never overlaps no effect
- EcoKI: never overlaps no effect
- EcoBI: never overlaps no effect
Digestion of Agarose-embedded DNA:
- Minimum 10U of the enzyme are required for complete digestion of 1µg of agarose-embedded lambda DNA in 4 hours
FspBI cleavage is impaired by overlappingdam methylation. To avoiddam methylation, use adam-, dcm strain such as GM2163 (#M0099).
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