OPTIZYME™ XbaI, Fisher BioReagents™
Brand: Fisher Bioreagents BP8008-5
Code : 32
Additional Details : Weight : 0.01000kg
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
- Buffer 1: 50 - 100%
- Buffer 2: 50 - 100%
- Buffer 3: 20 - 50%
- Buffer 4: 100%
- Buffer 5: 0 - 20%
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.
- XbaI is blocked by overlapping dam methylation.
- To avoid dam methylation, use a dam-, dcm- strain.
Dam: May overlap - blocked
- 5'...TCTAGm6A TC...3'
- 3'...AGATC Tm6AG...5'
- (Cleavage is blocked)
CpG: Never overlaps - no effect
EcoKI: Never overlaps - no effect
EcoBI: Never overlaps - no effect
|10X OPTIZYME™ Buffer 4|
|Indicates ligation efficiency of DNA fragments generated by digestion with the Restriction Enzyme|
|10mM Tris HCl (pH 7.4), 100mM KCl, 7mM 2-mercaptoethano, 1mM EDTA, 0.2mg/mL BSA, 50% Glycerol|