Recently Viewed

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

FspAI

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

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

The FspAI restriction enzyme recognizes RTGC^GCAY sites and cuts best at 37°C in O buffer
  
Loading...
Lambda DNA digested with FspAI

Lambda DNA digested with FspAI, 0.5% agarose, 2 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
Conditions for 100% Activity
  • 1X Buffer O: 50 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 100 mM NaCl, and 0.1 mg/mL BSA
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
NoteLow salt, high glycerol (>5%) concentrations, pH >8.0 or a large excess of enzyme may result in star activity.
Storage BufferFspAI 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
O (Orange) 37°C 0-20 0-20 100 50-100 0-20 50-100 2X

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: completely overlaps - blocked.
  • EcoKI: may overlap - effect not determined.
  • EcoBI: may overlap - effect not determined.
Methylation type Sequence Cleavage effect
CpGRTGCGCAY 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 20-50 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
2 0 0 2 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
0 0 0 0 0 2

New sites generated by ligation

Newly generated recognition sites resulting from ligation of blunt DNA ends

Recognition sequence Second restriction enzyme Restriction enzymes cleaving the newly generated recognition sequence
ATGC^GCAY
  • DraI/FastDigest DraI (TTT^AAA)
  • HincII (HindII)/FastDigest HincII* (GTY^AAC)
  • KspAI (HpaI)/FastDigest HpaI (KspAI) (GTT^AAC)
  • MssI (PmeI)/FastDigest MssI (GTTT^AAAC)
  • RsaI/FastDigest RsaI (GT^AC)
  • ScaI/FastDigest ScaI (AGT^ACT)
  • SmiI (SwaI)/FastDigest SwaI (SmiI) (ATTT^AAAT)
  • HpyCH4VI
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (CCG^CTC)
  • MnlI/FastDigest MnlI
  • Eco32I (EcoRV)/FastDigest EcoRV (Eco32I) (GAT^ATC)
  • LweI (SfaNI)/FastDigest SfaNI (BmsI)
  • HpyCH4V
  • Mph1103I (NsiI)/FastDigest NsiI (Mph1103I)
  • Eco47III (AfeI)/FastDigest AfeI (Eco47III) (AGC^GCT)
  • EheI (SfoI)/FastDigest EheI (GGC^GCC)
  • HhaI/FastDigest HhaI
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (GAG^CGG)
  • MspA1I* (CMG^CGG)
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • NsbI (FspI)/FastDigest FspI (NsbI) (TGC^GCA)
  • HhaI/FastDigest HhaI
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)
  • NsbI (FspI)/FastDigest FspI (NsbI)
  • PdiI (NaeI)/FastDigest NaeI (PdiI) (GCC^GGC)
  • SatI (Fnu4HI)/FastDigest Fnu4HI (SatI)
  • TauI/FastDigest TauI
  • SrfI (GCCC^GGGC)
  • Cac8I
  • SspI/FastDigest SspI (AAT^ATT)
  • HpyCH4V
  • Mph1103I (NsiI)/FastDigest NsiI (Mph1103I)
GTGC^GCAY
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI) (GG^CC)
  • CviJI* (RG^CC)
  • Eco147I (StuI)/FastDigest StuI (Eco147I) (AGG^CCT)
  • MlsI (MscI)/FastDigest MscI (MlsI) (TGG^CCA)
  • BseSI (Bme1580I)/FastDigest Bme1580I (BseSI)
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
  • DpnI/FastDigest DpnI (GA^TC)
  • Alw21I (BsiHKAI)/FastDigest Alw21I
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
  • DraI/FastDigest DraI (TTT^AAA)
  • HincII (HindII)/FastDigest HincII* (GTY^AAC)
  • KspAI (HpaI)/FastDigest HpaI (KspAI) (GTT^AAC)
  • MssI (PmeI)/FastDigest MssI (GTTT^AAAC)
  • SmiI (SwaI)/FastDigest SwaI (SmiI) (ATTT^AAAT)
  • SspI/FastDigest SspI (AAT^ATT)
  • HpyCH4V
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (CCG^CTC)
  • MnlI/FastDigest MnlI
  • Eco32I (EcoRV)/FastDigest EcoRV (Eco32I) (GAT^ATC)
  • LweI (SfaNI)/FastDigest SfaNI (BmsI)
  • HpyCH4V
  • Eco47III (AfeI)/FastDigest AfeI (Eco47III) (AGC^GCT)
  • EheI (SfoI)/FastDigest EheI (GGC^GCC)
  • HhaI/FastDigest HhaI
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (GAG^CGG)
  • MspA1I* (CMG^CGG)
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • NsbI (FspI)/FastDigest FspI (NsbI) (TGC^GCA)
  • HhaI/FastDigest HhaI
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)
  • NsbI (FspI)/FastDigest FspI (NsbI)
  • PdiI (NaeI)/FastDigest NaeI (PdiI) (GCC^GGC)
  • SatI (Fnu4HI)/FastDigest Fnu4HI (SatI)
  • TauI/FastDigest TauI
  • RsaI/FastDigest RsaI (GT^AC)
  • ScaI/FastDigest ScaI (AGT^ACT)
  • Alw21I (BsiHKAI)/FastDigest Alw21I
  • Alw44I (ApaLI)/FastDigest ApaLI (AIw44I)
  • BseSI (Bme1580I)/FastDigest Bme1580I (BseSI)
  • Hpy8I (MjaIV)/FastDigest Hpy8I
  • HpyCH4V
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
  • SrfI (GCCC^GGGC)
  • Cac8I