OPTIZYME™ XbaI, Fisher BioReagents™

Description

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

Supplied with: 10X OPTIZYME Buffer 4
Conditions for 100% Activity

• 1X OPTIZYME Buffer 4: 33mM Tris-acetate (pH 7.9 at 37°C), 10mM Mg-acetate, 66mM K-acetate and 0.1mg/ml BSA
• Incubate at 37°C

Enzyme Activity in OPTIZYME buffers:

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

Storage Buffer:
10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 7mM 2-mercaptoethanol, 1mM EDTA, 0.2mg/ml BSA and 50% (v/v) glycerol
Ligation and Re-cleavage:
More than 95% of the DNA fragments can be ligated and re-cut after a 50-fold over-digestion with XbaI.
Digestion of Agarose-embedded DNA:
A minimum 5 units of XbaI is required for the complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours.
Notes:

• XbaI is blocked by overlapping dam methylation.
• To avoid dam methylation, use a dam-, dcm- strain.

Compatible Ends: BcuI, Eco130I, NheI, XmaJI
Methylation Effects:
Dam: May overlap - blocked

• 5'...TCTAGm6A TC...3'
• 3'...AGATC Tm6AG...5'
• (Cleavage is blocked)

Dcm: Never overlaps - no effect
CpG: Never overlaps - no effect
EcoKI: Never overlaps - no effect
EcoBI: Never overlaps - no effect