Thermo Scientific™ Eco47III (AfeI)
The Eco47III (AfeI) restriction enzyme recognizes AGC^GCT sites and cuts best at 37°C in O buffer (Isoschizomers: AfeI, Aor51HI).
Manufacturer: Thermo Scientific™ ER0322
Lambda DNA digested with Eco47III (AfeI), 0.7% agarose, 2 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'...A G C▵G C T...3'
3'...T C G▵C G A...5'
Conditions for 100% Activity:
- 1X Buffer O:50mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl2, 100mM NaCl and 0.1mg/mL BSA
- Incubate at 37°C
- Eco47III is supplied in: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 Recleavage:
- After 50-fold overdigestion with Eco47III, more than 80% of pBR322 DNA fragments can be ligated and more than 90% of these can be recut
- Dam: never overlaps-no effect
- Dcm: never overlaps-no effect
- CpG: completely overlaps-blocked
- EcoKI: never overlaps-no effect
- EcoBI: may overlap-effect not determined
Digestion of Agarose-embedded DNA:
- Minimum 5U of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours