Thermo Scientific™ Cfr9I (XmaI) (10 U/µL) Thermo Scientific™ ER0172

Description

The Cfr9I (XmaI) restriction enzyme recognizes C^CCGGG sites and cuts best at 37°C in its own unique buffer (Isoschizomers: TspMI, XmaCI, XmaI).

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

Storage Buffer:

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

Methylation Effects:

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

Note:

To achieve complete digestion of substrate with Cfr9I, the concentration of DNA should be no less than 50μg/mL in the reaction buffer.