Recently Viewed

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

TatI

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

FastDigest enzyme available Use Tango buffer for 100% activity Optimal incubation at 65°C Star activity Not inactivated at 80°C in 20 min. LO certified

The TatI restriction enzyme recognizes W^GTACW sites and cuts best at 65°C in Tango buffer
  
Loading...
Lambda DNA digested with TatI

Lambda DNA digested with TatI, 1% agarose, 24 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 EndsAcc65I, BshNI, Bsp1407I, Pfl23II.
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 65°C
  • To ensure higher efficiency of digestion, perform the cleavage reaction under paraffin oil
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
NoteIncubation at 37°C results in 20% activity. Greater than 5-fold overdigestion with TatI may result in star activity.
Storage BufferTatI is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 100 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 65°C NR 50-100† 0-20 20-50 100† 0-20 1X†

† Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).

Methylation Effects

  • Dam: never overlaps – no effect.
  • Dcm: never overlaps – no effect.
  • CpG: never overlaps – no effect.
  • EcoKI: never overlaps – no effect.
  • EcoBI: may overlap – effect not determined.

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
0-20 20-50 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
24 0 5 2 2 2
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
1 1 1 1 1 1

New sites generated by ligation

Newly generated recognition sites resulting from ligation of protruding compatible DNA ends

Recognition SequenceSecond EnzymeEnzymes recognizing newly generated recognition sequence
A^GTACW
  • Acc65I (Asp718I)/FastDigest Acc65I (G^GTACC)
  • BshNI (BanI)/FastDigest BanI (BshNI)* (G^GTACC)
  • Pfl23II (BsiWI)/FastDigest BsiWI (Pfl23II) (C^GTACG)
  • Csp6I (CviQI)/FastDigest Csp6I
  • RsaI/FastDigest RsaI
  • Bsp1407I (BsrGI)/FastDigest Bsp1407I (T^GTACA)
  • Csp6I (CviQI)/FastDigest Csp6I
  • RsaI/FastDigest RsaI
  • TatI/FastDigest TatI
T^GTACW
  • Acc65I (Asp718I)/FastDigest Acc65I (G^GTACC)
  • BshNI (BanI)/FastDigest BanI (BshNI)* (G^GTACC)
  • Pfl23II (BsiWI)/FastDigest BsiWI (Pfl23II) (C^GTACG)
  • Csp6I (CviQI)/FastDigest Csp6I
  • RsaI/FastDigest RsaI
  • Bsp1407I (BsrGI)/FastDigest Bsp1407I (T^GTACA)
  • Bsp1407I (BsrGI)/FastDigest Bsp1407I
  • Csp6I (CviQI)/FastDigest Csp6I
  • RsaI/FastDigest RsaI
  • TatI/FastDigest TatI

Newly generated recognition sites resulting from fill-in of 5'-overhang and self-ligation

Recognition SequenceNewly generated sequence after reactionEnzymes that cleave the newly generated sequence
W^GTACWWGTACGTACW
  • [Csp6I (CviQI)/FastDigest Csp6I]
  • Eco105I (SnaBI)/FastDigest SnaBI (Eco105I)
  • MaeII
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)
  • [RsaI/FastDigest RsaI]
  • SetI
  • TaiI (MaeII)/FastDigest TaiI