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HphI

5'...G G T G A (N)8↓...3'
3'...C C A C T (N)7↑...5

B buffer for 100% activity Optimal incubation at 37°C Cleavage blocked or impaired by overlapping Dam methylation Thermal inactivation at 65°C in 20 min Recombinant enzyme LO certified

The HphI restriction enzyme recognizes GGTGA(8/7)^ sites and cuts best at 37°C in B buffer (Isoschizomers: AsuHPI)
  
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Lambda DNA (dam-) digested with HphI

Lambda DNA (dam- digested with HphI, 1.4% agarose, 168 cleavage sites

Thermo Scientific 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.

Highlights

  • Superior quality – stringent quality control and industry leading manufacturing process
  • Convenient color-coded Five Buffer System
  • Includes universal Tango buffer for double-digestions
  • BSA premixed in reaction buffers
  • Wide selection of restriction endonuclease specificities

Applications

  • Molecular cloning
  • Restriction site mapping
  • Genotyping
  • Southern Blot
  • Restriction fragment length polymorphism (RFLP)
  • SNP

Note

Assayed using lambda DNA (dam-) (#SD0021). HphI is blocked by overlapping dam methylation. To avoid dam methylation, use a dam-, dcm- strain, such as GM2163. High glycerol (> 5%) concentrations, pH > 8.0 or a large excess of enzyme may result in star activity.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • 1X Buffer B: 10 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, and 0.1 mg/mL BSA
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferHphI is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/mL BSA, and 50% (v/v) glycerol.

Reaction conditions

Recommended buffer for 100% activity Optimal temperature Enzyme activity in Thermo Scientific buffers, % Tango buffer for double digestion
B (blue)
1X		 G (green)
1X		 O (orange)
1X		 R (red)
1X		 Tango
(yellow)
1X / 2X
B (Blue) 37°C 100 0-20 0-20 0-20 20-50 0-20 1X

Methylation Effects

Methylation type Sequence Cleavage effect
Dam (GATC) 5'...GGTGm6A TC...3'
3'...CCAC Tm6AG...5'		 Blocked
Dcm (CCWGG) 5'...Cm5CW GGTGA...3'
3'...G GWm5CCACT...5'		 No effect
CpG 5'...m5C GGTGA...3'
3'... Gm5CCACT...5'		 No effect
EcoBI (TGA(N)8TGCT) 5'...GGTGm6A(N)8 TGCT...3'
3'...CCAC T(N)8m6ACGA...5'		 Blocked

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
0 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
168 9 18 12 7 7
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
6 6 6 6 17 16

 

MSDS & Certificates

pdf   HphI - MSDS
pdf   MSDS (JP) - Restriction Enzymes