Thermo Scientific™ HphI (10 U/μL)

Description

The HphI restriction enzyme recognizes GGTGA(8/7)^ sites and cuts best at 37°C in B buffer (Isoschizomers: AsuHPI).

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

Storage Buffer:

• 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

Methylation Effects:

• Dam: may overlap —blocked
• Dcm: may overlap —no effect
• CpG: may overlap —no effect
• EcoKI: never overlaps —no effect
• EcoBI: may overlap —blocked

Note:

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.

Specifications

• Concentration: 10 U/μL
• Methylation Sensitivity: Dam Methylation-Sensitive, Not CpG Methylation-Sensitive, Not Dcm Methylation-Sensitive
• Enzyme: HphI
• Compatible Buffer: 10X Buffer B
• Sensitive to Heat Inactivation: Yes
• Optimal Reaction Temperature: 37°C
• Type IIS RE: Yes
• Quantity: 300 units
• Product Type: Restriction Enzyme
• Research Category: Scarless Cloning

For Research Use Only. Not for use in diagnostic procedures.