Thermo Scientific™ SatI (Fnu4HI)
The SatI (Fnu4HI) restriction enzyme recognizes GC^NGC sites and cuts best at 37°C in G buffer (Isoschizomers: Fnu4HI, Fsp4HI, ItaI).
Manufacturer: Thermo Scientific™ ER1642
Lambda DNA digested with SatI (Fnu4HI), 1.4% agarose, 380 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.5'...G C▵N G C...3'
3'...C G N▵C G...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
- SatI 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 50-fold overdigestion with SatI, more than 60% of the DNA fragments can be ligated in a reaction mixture containing 10-30U of T4 DNA Ligase/1μg of fragments and 10% PEG
- More than 90% of these can be recut
- Dam: never overlaps — no effect
- Dcm: never overlaps — no effect
- CpG: may overlap — blocked
- EcoKI: never overlaps — no effect
- EcoBI: never overlaps — no effect
- GC^SGC — BcnI, Bme1390I
- GC^WGC — MvaI, Bme1390I
At least two copies of SatI (Fnu4HI) recognition site are required for efficient cleavage.