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Eco32I (EcoRV)

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

FastDigest enzyme available R buffer for 100% activity Optimal incubation at 37°C Thermal inactivation at 80°C in 20 min High concentration available Genome qualified LO certified

The Eco32I (EcoRV) restriction enzyme recognizes GAT^ATC sites and cuts best at 37°C in R buffer (Isoschizomers: EcoRV)
  
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Lambda DNA digested with Eco32I (EcoRV)

Lambda DNA digested with Eco32I (EcoRV), 1% agarose, 21 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 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 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 BufferEco32I is supplied in: 25 mM Tris-HCl (pH 7.5 at 25°C), 200 mM NaCl, 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) 37°C 0-20 50-100 50-100 100 20-50 100 1X or 2X

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: may overlap - no effect.
  • EcoKI: never overlaps - no effect.
  • EcoBI: may overlap - effect not determined.
Methylation type Sequence Cleavage effect
CpG 5'...m5C GATATm5C G...3'
3'... Gm5CTATA Gm5C...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
21 0 0 1 0 1
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
0 1 1 1 0 1

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
GAT^ATC
  • AanI (PsiI)/FastDigest PsiI (AanI) (TTA^TAA)
  • FastDigest MseI (SaqAI)
  • Tru1I (MseI)/FastDigest Tru1I
  • AluI/FastDigest AluI (AG^CT)
  • Bsh1236I (BstUI)/FastDigest Bsh1236I (CG^CG)
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI) (GG^CC)
  • CviJI* (RG^CC)
  • CviJI* (RG^CT)
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • Eco147I (StuI)/FastDigest StuI (Eco147I) (AGG^CCT)
  • HpyCH4V (TG^CA)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (CCG^CTC)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (GAG^CGG)
  • MlsI (MscI)/FastDigest MscI (MlsI) (TGG^CCA)
  • MspA1I* (CMG^CGG)
  • MspA1I* (CMG^CTG)
  • PvuII/FastDigest PvuII (CAG^CTG)
  • Bsp143I (Sau3AI)/FastDigest Sau3AI (Bsp143I)
  • DpnI/FastDigest DpnI
  • MboI/FastDigest MboI
  • Bsp68I (NruI)/FastDigest NruI (RruI) (TCG^CGA)
  • Bsp143I (Sau3AI)/FastDigest Sau3AI (Bsp143I)
  • DpnI/FastDigest DpnI
  • MboI/FastDigest MboI
  • TaqI/FastDigest TaqI
  • DpnI/FastDigest DpnI (GA^TC)
  • HinfI/FastDigest HinfI
  • PfeI (TfiI)/FastDigest TfiI (PfeI)
  • FspAI/FastDigest FspAI* (RTGC^GCAC)
  • NsbI (FspI)/FastDigest FspI (NsbI) (TGC^GCA)
  • HpyCH4V
  • FspAI/FastDigest FspAI* (RTGC^GCAT)
  • HpyCH4V
  • Mph1103I (NsiI)/FastDigest NsiI (Mph1103I)