Thermo Scientific™ Cfr9I (XmaI)
The Cfr9I (XmaI) restriction enzyme recognizes C^CCGGG sites and cuts best at 37°C in its own unique buffer (Isoschizomers: TspMI, XmaCI, XmaI).
Manufacturer: Thermo Scientific™ ER0172
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'...C▵C C G G G...3'
3'...G G G C C▵C...5'
Conditions for 100% Activity:
- 1X Buffer Cfr9I:10mM Tris-HCl (pH 7.2 at 37°C), 5mM MgCl2, 200mM sodium glutamate and 0.1mg/mL BSA
- Incubate at 37°C
- Cfr9I is supplied in:10mM Tris-HCl (pH 7.5 at 25°C), 250mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol
Ligation and Recleavage:
- After 50-fold overdigestion with Cfr9I, more than 95% of the DNA fragments can be ligated and recut
- Dam: never overlaps-no effect
- Dcm: never overlaps-no effect
- CpG: completely overlaps-cleavage impaired
- EcoKI: never overlaps-no effect
- EcoBI: never overlaps-no effect
Digestion of Agarose-embedded DNA:
- Minimum 5U of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours
To achieve complete digestion of substrate with Cfr9I, the concentration of DNA should be no less than 50μg/mL in the reaction buffer.