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FastDigest Tsp509I (TasI)

5'...↓A A T T ...3'
3'... T T A A↑...5'

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

The FastDigest Tsp509I (TasI) restriction enzyme recognizes ^AATT sites and cuts best at 65°C in 5-15 minutes using universal Yellow Tango Buffer (Isoschizomers: Sse9I, TspEI)
  
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Lambda DNA digested with HphI

pBR322 DNA, 1.0% agarose, 8 cleavage sites

Thermo Scientific FastDigest enzymes are an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.

The universal buffer allows rapid single-, double- or multiple DNA digestion within 5-15 minutes eliminating any need for buffer change or subsequent DNA clean-up steps. DNA modifying enzymes, such as Klenow Fragment, T4 DNA Ligase, alkaline phosphatases and T4 DNA Polymerase all have 100% activity in FastDigest Buffer. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.

For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels.

Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects.

Features

  • 100% activity of all FastDigest enzymes in the universal buffer
  • 100% buffer compatibility with downstream applications
  • Complete digestion in 5-15 minutes
  • Direct loading on gels
  • No star activity
  • 176 FastDigest enzymes available

Applications

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

Note

The FastDigest Green Buffer offers the same high performance in DNA digestion and downstream applications as the colorless FastDigest Buffer. For applications that require product analysis by fluorescence excitation (e.g. concentration measurements in UV light) the colorless FastDigest Buffer is recommended.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Compatible EndsFastDigest EcoRI, FastDigest MfeI (MunI), FastDigest XapI, EcoRI, MunI, XapI.
Formulation1 μL of enzyme (1 FDU) cleaves 1 μg of pBR322 DNA in 5 minutes at 65°C in 1X FastDigest Buffer.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Recommended Reaction Conditions

1X FastDigest buffer or 1X FastDigest Green buffer at 65°C.
1 µL of FastDigest Tsp509I (TasI) is formulated to digest up to:

  • 1 µg of pBR322 DNA in 5 minutes
  • 1 µg of plasmid DNA in 5 minutes
  • 0.2 µg of PCR product in 5 minutes
  • 1 µg of genomic DNA in 5 minutes or 5 µg of genomic DNA in 30 minutes

Reaction conditions

Reaction temperature Digestion time with 1 µl of FastDigest enzyme, min bp from end of DNA required for complete digestion Thermal inactivation Incubation time without star activity, hours
Lambda, 1 µg/20 µl Plasmid DNA, 1 µg/20 µl PCR product, ~0.2 µg/30 µl Genomic DNA, 1 µg/10 µl
65°C 5
(pBR322 DNA)		 5 5 5 3 No
(chloroform extraction)		 6

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

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)