Thermo Scientific™ TaaI (HpyCH4III)
The TaaI (HpyCH4III) restriction enzyme recognizes ACN^GT sites and cuts best at 65°C in Tango buffer (Isoschizomers: Bst4CI, HpyCH4III).
Manufacturer: Thermo Scientific™ ER1362
Lambda DNA digested with TaaI (HpyCH4III), 1.4% agarose, 187 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 C N▵G T...3'
3'...T G▵N C A...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 65°C
- To ensure higher efficiency of digestion, perform the cleavage reaction under paraffin oil
- TaaI is supplied in:10mM Tris-HCl (pH7.5 at 25°C), 100mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol
Ligation and Recleavage
- After 50-fold overdigestion with TaaI, approximately 90% of the DNA fragments can be ligated and recut
- Dam: never overlaps — no effect
- Dcm: never overlaps — no effect
- CpG: may overlap — cleavage impaired
- EcoKI: may overlap — effect not determined
- EcoBI: may overlap — effect not determined
Incubation at 37°C results in 10% activity.