Thermo Scientific™ SspI (10 U/μL)

Description

Lambda DNA digested with SspI, 1% agarose, 20 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 T A▵T A A...5'

Conditions for 100% Activity

• 1X Buffer G:10mM Tris-HCl (pH7.5 at 37°C), 10mM MgCl₂, 50mM NaCl and 0.1mg/mL BSA.Incubate at 37°C

Storage Buffer

• SspI is supplied in:10mM Tris-HCl (pH7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol

Ligation and Recleavage

• After 10-fold overdigestion with SspI, more than 90% of the DNA fragments can be ligated and 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 — no effect

Digestion of Agarose-embedded DNA

• Minimum 5units of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16hours

Note:

Greater than 10-fold overdigestion with SspI may result in star activity.