OPTIZYME™ XhoI, Fisher BioReagents™

5'....C^T C G A G...3'
3'....G A G C T^C...5'

Supplied with: 10X OPTIZYME Buffer 5
Conditions for 100% Activity:

• 1X OPTIZYME Buffer 5: 10mM Tris-HCl (pH 8.5 at 37°C), 10mM MgCl₂ , 100mM KCl and 0.1mg/ml BSA
• Incubate at 37°C

• Buffer 1: 0 - 20%
• Buffer 2: 50 - 100%
• Buffer 3: 50 - 100%
• Buffer 4: 20 - 50%
• Buffer 5: 100%

• 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 Re-cleavage:
• More than 95% of DNA fragments can be ligated and re-cut after a 50-fold over-digestion with XhoI.
• Digestion of Agarose-embedded DNA:
• A minimum of 5 units of XhoI is required for the complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours.
• Note:
• Supercoiled plasmids may require up to 5-fold more XhoI for complete digestion than linear DNA.
• Compatible Ends: BauI, Eco88I, SalI, SmoI
• Methylation Effects:
• Dam: Never overlaps - no effect
• Dcm: Never overlaps - no effect
• CpG: Completely overlaps - cleavage impaired
• CTCGAG

(Cleavage impaired)
EcoKI: Never overlaps - no effect
EcoBI: Never overlaps - no effect