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DpnI

5'...G m6A  T C...3'
3'...C   Tm6A G...5'

Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available Tango buffer for 100% activity Optimal incubation at 37°C Thermal inactivation at 80°C in 20 min Recombinant enzyme LO certified

The DpnI restriction enzyme recognizes Gm6A^TC sites and cuts best at 37°C in Tango buffer (Isoschizomers: MalI)
  
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pBR322 DNA digested with DpnI

pBR322 DNA digested with DpnI, 1.4% agarose, 22 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

DpnI requires the presence of N6-methyladenine within the recognition sequence to cleave DNA. DNA purified from a dam+ strain will be a substrate for DpnI. DpnI will only cleave fully-adenomethylated dam sites. Hemi-adenomethylated dam sites DpnI cleaves 60X more slowly. DpnI, Bsp143I, and MboI all recognize the same sequence, but have different methylation sensitivities and cleavage sites. Assayed using pBR322 DNA.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • 1X Buffer Tango: 33 mM Tris-acetate (pH 7.9 at 37°C), 10 mM Mg-acetate, 66 mM K-acetate, and 0.1 mg/mL BSA
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferDpnI is supplied in: 10 mM Tris-HCl (pH 7.4 at 25°C), 400 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
Tango 37°C 100 100 50-100 50-100 100 50-100 1X or 2X

Methylation Effects

Methylation type Sequence Cleavage effect
CpG 5'...Gm6A Tm5C G...3'
3'...C Tm6A Gm5C...5'
Cleaves only Dam methylated DNA
Dam (GATC)Gm6ATC Cleaves only Dam methylated DNA

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
116 0 7 22 15 15
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
15 15 15 15 22 15

New sites generated by ligation

Newly generated recognition sites resulting from ligation of blunt DNA ends

Recognition sequence Second restriction enzyme Restriction enzymes cleaving the newly generated recognition sequence
GA^TC
  • AjiI (BmgBI)* (CAC^GTC)
  • ZraI (GAC^GTC)
  • HinfI/FastDigest HinfI
  • PleI
  • SchI (MlyI)/FastDigest MlyI (SchI)
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (CCG^CTC)
  • HinfI/FastDigest HinfI
  • Eco147I (StuI)/FastDigest StuI (Eco147I) (AGG^CCT)
  • SetI
  • Eco32I (EcoRV)/FastDigest EcoRV (Eco32I) (GAT^ATC)
  • HinfI/FastDigest HinfI
  • PfeI (TfiI)/FastDigest TfiI (PfeI)
  • Eco47III (AfeI)/FastDigest AfeI (Eco47III) (AGC^GCT)
  • AluI/FastDigest AluI
  • CviJI
  • SetI
  • EheI (SfoI)/FastDigest EheI (GGC^GCC)
  • CviJI
  • FspAI/FastDigest FspAI* (RTGC^GCAC)
  • Alw21I (BsiHKAI)/FastDigest Alw21I
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
  • SspI/FastDigest SspI (AAT^ATT)
  • TasI (Tsp509I)/FastDigest Tsp509I (TasI)