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TasI (Tsp509I)

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

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

The TasI (Tsp509I) restriction enzyme recognizes ^AATT sites and cuts best at 65°C in B buffer (Isoschizomers: Sse9I, Tsp509I, TspEI)
  
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Lambda DNA digested with TasI (Tsp509I)

Lambda DNA digested with TasI (Tsp509I), 1% agarose, 189 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 EndsEcoRI, MunI, XapI.
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 65°C
  • To ensure higher efficiency of digestion, perform the cleavage reaction under paraffin oil
Double DigestionPerform double digestion using DoubleDigest.
Isoschizomers Search for commercial isoschizomers using REsearch.
NoteIncubation at 37°C results in less than 10% activity.
Storage BufferTasI 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) 65°C 100 50-100 20-50 0-20 20-50 0-20 1X

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 - blocked.
Methylation type Sequence Cleavage effect
EcoBI (TGA(N)8TGCT) 5'...TGm6AATT(N)5 TGCT...3'
3'...AC TTAA(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
0 20-50 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
189 25 64 8 7 7
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
10 10 12 12 14 13

New sites generated by ligation

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

Recognition Sequence Second Enzyme Enzymes recognizing newly generated recognition sequence
^AATT
  • EcoRI/FastDigest EcoRI (G^AATTC)
  • MunI (MfeI)/FastDigest MfeI (MunI) (C^AATTG)
  • XapI (ApoI)/FastDigest XapI* (R^AATTC)
  • XapI (ApoI)/FastDigest XapI* (R^AATTT)
  • TasI (Tsp509I)/FastDigest Tsp509I (TasI)

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

Recognition Sequence Newly generated sequence after reaction Enzymes that cleave the newly generated sequence
^AATTAATTAATT
  • FastDigest MseI (SaqAI)
  • [TasI (Tsp509I)/FastDigest Tsp509I (TasI)]
  • Tru1I (MseI)/FastDigest Tru1I
  • VspI (AseI)/FastDigest AseI (VspI)