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Bsu15I (ClaI)

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

FastDigest enzyme available Tango buffer for 100% activity Cleavage blocked or impaired by CpG methylation Cleavage blocked or impaired by overlapping Dam methylation Optimal incubation at 37°C Thermal inactivation at 65°C in 20 min Genome qualified LO certified

The Bsu15I (ClaI) restriction enzyme recognizes AT^CGAT sites and cuts best at 37°C in Tango buffer (Isoschizomers: BanIII, Bsa29I, BseCI, BshVI, BspDI, BspXI, BsuTUI, ClaI)
  
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Lambda DNA digested with Bsu15I (ClaI)

Lambda DNA digested with Bsu15I (ClaI), 0.7% agarose, 15 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

Bsu15I is blocked by overlapping dam methylation. To avoid dam methylation, use a dam-, dcm- strain, such as GM2163.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Compatible EndsBsp119I, Hin1I, Hin6I, HpaII, MaeII, MspI, NarI, Psp1406I, SsiI, TaqI, XmiI.
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
Digestion of Agarose-embedded DNAMinimum 5 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferBsu15I is supplied in: 10 mM Tris-HCl (pH 8.0 at 25°C), 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.5 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 20-50 20-50 20-50 20-50 100 20-50 1X or 2X

Methylation Effects

Methylation type Sequence Cleavage effect
Dam (GATC) 5'...ATCGm6A TC...3' 3'...TAGC Tm6AG...5' Blocked
CpGATCGAT Blocked

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
15 0 2 1 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 1 1 1 1

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
AT^CGAT
  • BmeT110I* (CY^CGAG)
  • Bsp119I (BstBI)/FastDigest Bsp119I (TT^CGAA)
  • TaqI/FastDigest TaqI (T^CGA)
  • XmiI (AccI)/FastDigest AccI (XmiI)* (GT^CGAC)
  • TaqI/FastDigest TaqI

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
AT^CGATATCGCGAT
  • Bsh1236I (BstUI)/FastDigest Bsh1236I
  • Hpy188III
  • Bsp68I (NruI)/FastDigest NruI (RruI)