Thermo Scientific™ ScaI (10 U/μL)

Description

Lambda DNA digested with ScaI, 0.7% agarose, 5 cleavage sites

conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

3'...T C A▵T G A...5'

Conditions for 100% Activity

• 1X Buffer ScaI:10mM Bis-Tris Propane-HCl (pH6.5 at 37°C), 10mM MgCl₂, 100mM KCl and 0.1mg/mL BSA
• Incubate at 37°C

Storage Buffer

• ScaI is supplied in:10mM Tris-HCl (pH7.5 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/mL BSA and 50% (v/v) glycerol

Ligation and Recleavage

• After 50-fold overdigestion with ScaI, more than 80% of the DNA fragments can be ligated and more than 90% of these can be recut

Methylation Effects

• Dam: never overlaps — no effect
• Dcm: never overlaps — no effect
• CpG: never overlaps — no effect
• EcoKI: never overlaps — no effect
• EcoBI: may overlap — blocked

Digestion of Agarose-embedded DNA

• Minimum 20units of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16hours

Note:

Conditions of low salt, high enzyme concentration, glycerol concentration > 5% or pH> 8.0 may result in star activity. Supercoiled plasmids may require up to 20-fold more ScaI for complete digestion than linear DNA.