OPTIZYME™ SmaI, Fisher BioReagents™

Description

5'...C C C^G G G...3'
3'...G G G^C C C...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 30°C

Enzyme Activity in OPTIZYME buffers:

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

Storage Buffer:

• 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 90% of the pUC19 DNA fragments can be ligated and re-cut after 50-fold over-digestion with SmaI.

Digestion of Agarose-embedded DNA:

A minimum of 5 units of SmaI is required for the complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours.

Notes:

• Incubation at 37°C results in 50% activity. SmaI has a half-life of 15 min at 37°C.
• Incubation at 25°C results in 100% activity.
• SmaI needs K+ to work for activity.

Methylation Effects:
Dam: Never overlaps - no effect
Dcm: Never overlaps - no effect
CpG: Completely overlaps - blocked

• CCCGGG

(Cleavage blocked)

EcoKI: Never overlaps - no effect

EcoBI: Never overlaps - no effect