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

FspBI (BfaI)

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

FastDigest enzyme available Tango buffer for 100% activity Optimal incubation at 37°C Thermal inactivation at 65°C in 20 min Genome qualified LO certified

The FspBI (BfaI) restriction enzyme recognizes C^TAG sites and cuts best at 37°C in Tango buffer (Isoschizomers: BfaI, MaeI, XspI)
  
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Lambda DNA digested with FspBI (BfaI)

Lambda DNA digested with FspBI (BfaI), 0.7% agarose, 13 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

FspBI cleavage is impaired 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 EndsCsp6I, NdeI, Tru1I, VspI.
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 10 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 4 hours.
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferFspBI 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
Tango 37°C 50-100 20-50 0-20 0-20 100 0-20 1X

Methylation Effects

  • Dam: may overlap - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: never overlaps - no effect.
  • EcoKI: never overlaps - no effect.
  • EcoBI: never overlaps - no effect.
Methylation type Sequence Cleavage effect
Dam (GATC) 5'...CTAGm6A TC...3'
3'...GATC Tm6AG...5'		 No effect

Cleavage efficiency close to the termini of PCR fragments

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

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
13 3 5 5 4 4
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
5 5 6 6 4 4

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
C^TAG
  • NdeI/FastDigest NdeI (CA^TATG)
  • FaiI

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
C^TAGCTATAG
  • BfmI (SfcI)/FastDigest SfcI (BfmI)
  • FaiI