Thermo Scientific™ HphI
The HphI restriction enzyme recognizes GGTGA(8/7)^ sites and cuts best at 37°C in B buffer (Isoschizomers: AsuHPI).
Manufacturer: Thermo Scientific™ ER1101
Lambda DNA (dam- digested with HphI, 1.4% agarose, 168 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 G T G A (N)8▵...3'
3'...C C A C T (N)7▵...5'
Conditions for 100% Activity:
- 1X Buffer B: 10mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl2 and 0.1mg/mL BSA
- Incubate at 37°C
- HphI is supplied in: 10mM Tris-HCl (pH 7.5 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol
- Dam: may overlap —blocked
- Dcm: may overlap —no effect
- CpG: may overlap —no effect
- EcoKI: never overlaps —no effect
- EcoBI: may overlap —blocked
Assayed using lambda DNA (dam) (#SD0021). HphI is blocked by overlappingdam methylation. To avoiddam methylation, use adam-, dcm strain such as GM2163 (#M0099). High glycerol (>5%) concentrations, pH>8.0 or a large excess of enzyme may result in star activity.