Thermo Scientific™ Eco105I (SnaBI)
The Eco105I (SnaBI) restriction enzyme recognizes TAC^GTA sites and cuts best at 37°C in Tango buffer (Isoschizomers: BstSNI, SnaBI).
Manufacturer: Thermo Scientific™ ER0402
Lambda DNA digested with Eco105I (SnaBI), 0.7% agarose, 1 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'...T A C▵G T A...3'
3'...A T G▵C A T...5'
Conditions for 100% Activity:
- 1X Buffer Tango™:
- 33mM Tris-acetate (pH7.9 at 37°C), 10mM Mg-acetate, 66mM K-acetate and 0.1mg/ml BSA.Incubate at 37°C
- Eco105I is supplied in:10mM Tris-HCl (pH7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 1mM phenylmethylsulfonylfluoride, 0.2mg/ml BSA and 50% (v/v) glycerol
Ligation and Recleavage:
- After 10-fold overdigestion with Eco105I, more than 80% of the M13mp18 DNA fragments can be ligated in a reaction mixture containing 20-40U 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: completely overlaps — blocked
- EcoKI: never overlaps — no effect
- EcoBI: never overlaps — 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
Greater than 15-fold overdigestion with Eco105I may result in star activity.