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

RsaI

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

FastDigest enzyme available Tango buffer for 100% activity Optimal incubation at 37°C Cleavage blocked or impaired by overlapping CpG methylation Thermal inactivation at 80°C in 20 min LO certified

The RsaI restriction enzyme recognizes GT^AC sites and cuts best at 37°C in Tango buffer (Isoschizomers: AfaI, CviQI, RsaNI)
  
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Lambda DNA digested with RsaI

Lambda DNA digested with RsaI, 1.4% agarose, 113 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 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
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferRsaI 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

Methylation type Sequence Cleavage effect
CpG

5'...GTAm5C G...3'
3'...CAT Gm5C...5'

 Impaired
CpG

5'...m5C GTAm5C G...3'
3'... Gm5CAT Gm5C...5'

 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
113 11 19 3 3 3
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
2 2 2 2 4 3

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
GT^AC
  • AanI (PsiI)/FastDigest PsiI (AanI) (TTA^TAA)
  • FastDigest MseI (SaqAI)
  • Tru1I (MseI)/FastDigest Tru1I
  • Bsp68I (NruI)/FastDigest NruI (RruI) (TCG^CGA)
  • TaqI/FastDigest TaqI
  • Bst1107I (BstZ17I)/FastDigest BstZ17I (Bst1107I) (GTA^TAC)
  • HincII (HindII)/FastDigest HincII* (GTY^AAC)
  • KspAI (HpaI)/FastDigest HpaI (KspAI) (GTT^AAC)
  • MaeIII
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (CCG^CTC)
  • Alw26I (BsmAI)/FastDigest Alw26I
  • FspAI/FastDigest FspAI* (RTGC^GCAC)
  • Alw21I (BsiHKAI)/FastDigest Alw21I
  • Alw44I (ApaLI)/FastDigest ApaLI (Alw44I)
  • BseSI (Bme1580I)/FastDigest Bme1580I (BseSI)
  • Hpy8I (MjaIV)/FastDigest Hpy8I
  • HpyCH4V
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
  • FspAI/FastDigest FspAI* (RTGC^GCAT)
  • NsbI (FspI)/FastDigest FspI (NsbI) (TGC^GCA)
  • HpyCH4V
  • HincII (HindII)/FastDigest HincII* (GTY^GAC)
  • MaeIII
  • NmuCI (Tsp45I)/FastDigest NmuCI
  • MssI (PmeI)/FastDigest MssI (GTTT^AAAC)
  • Hpy8I (MjaIV)/FastDigest Hpy8I
  • ScaI/FastDigest ScaI (AGT^ACT)
  • Csp6I (CviQI)/FastDigest Csp6I
  • RsaI/FastDigest RsaI

MSDS & Certificates

pdf   RsaI - MSDS
pdf   MSDS (JP) - Restriction Enzymes