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Home | Molecular Biology | Restriction & Modifying Enzymes | Restriction Enzymes | Conventional

HinfI

5'...G↓A N T C...3'
3'...C T N A↑G...5'
Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available R buffer for 100% activity Optimal incubation at 37°C Cleavage blocked or impaired by CG methylation Thermal inactivation at 65°C for 20 min. Available in high concentration (50 U/µL) LO certified

The HinfI restriction enzyme recognizes G^ANTC sites and cuts best at 37°C in R buffer
  
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Lambda DNA digested with HinfI

Lambda DNA digested with HinfI, 1.4% agarose, 148 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

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Compatible EndsG↓AWTC – PfeI.
Conditions for 100% Activity
  • 1X Buffer R: 10 mM Tris-HCl (pH 8.5 at 37°C), 10 mM MgCl2, 100 mM KCl, and 0.1 mg/mL BSA
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferHinfI is supplied in: 10 mM Tris-HCl (pH 7.4 at 25°C), 100 mM KCl, 1 mM DTT, 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
R (Red) 37°C 0-20 20-50 50-100 100 50-100 50-100 1X or 2X

Methylation Effects

Methylation type Sequence Cleavage effect
CpG 5'...GANTm5C G...3'
3'...CTNA Gm5C...5'		 Impaired
CpG 5'...m5C GANTm5C G...3'
3'... Gm5CTNA Gm5C...5'		 Impaired
EcoBI (TGA(N)8TGCT) 5'...TGm6ANTC(N)5 TGCT...3'
3'...AC TNAG(N)5m6ACGA...5'		 Blocked

Cleavage efficiency close to the termini of PCR fragments

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

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
148 21 27 10 6 5
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
9 8 8 8 13 9