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Tru1I (MseI)

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

FastDigest enzyme available Use Red buffer for 100% activity Optimal incubation at 65°C Not inactivated at 80°C in 20 min. Available in high concentration (50 U/µL) LO Certified

The Tru1I (MseI) restriction enzyme recognizes T^TAA sites and cuts best at 65°C in R buffer (Isoschizomers: MseI, Tru9I)
  
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Lambda DNA digested with TrulI (MseI)

Lambda DNA digested with TrulI (MseI), 1.4% agarose, 195 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 EndsCsp6I, FspBI, NdeI, VspI.
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 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 10% activity. We recommend using Tru1I, HC at 37°C.
Storage BufferTru1I 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) 65°C 50-100 50-100 20-50 100 50-100 100 1X or 2X

Methylation Effects

  • Dam: never overlaps – no effect.
  • Dcm: never overlaps – no effect.
  • CpG: never overlaps – no effect.
  • EcoKI: may overlap – blocked.
  • EcoBI: never overlaps – no effect.
Methylation type Sequence Cleavage effect
EcoKI(AAC(N)6GTGC) 5'...TTAm6AC(N)6G  TGC...3'
3'...AAT  TG(N)6Cm6ACG...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 0-20 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
195 35 63 15 13 13
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
18 18 19 19 16 12

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
T^TAA
  • NdeI/FastDigest NdeI (CA^TATG)
  • FaiI
  • FastDigest MseI (SaqAI) (T^TAA)
  • VspI (AseI)/FastDigest AseI (VspI) (AT^TAAT)
  • FastDigest MseI (SaqAI)
  • Tru1I (MseI)/FastDigest Tru1I

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
T^TAATTATAA
  • AanI (PsiI)/FastDigest PsiI (AanI)
  • FaiI