5'...G↓A A T T C...3'
3'...C T T A A↑G...5'
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.
- 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
- Molecular cloning
- Restriction site mapping
- Southern Blot
- Restriction fragment length polymorphism (RFLP)
For methylation sensitivity refer to product specifications.
|Compatible Ends||XapI, MunI, TasI.|
|Conditions for 100% Activity||
|Digestion of Agarose-embedded DNA||Minimum 5 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.|
|Double Digestion||Perform double digestion using DoubleDigest.|
|Isoschizomers||Search for commercial isoschizomers using REsearch.|
|Note||Low salt concentration, large excess of the enzyme, pH > 8.0, or the replacement of Mg2+ by Mn2+ may result in star activity.|
|Storage Buffer||EcoRI is supplied in: 10 mM potassium phosphate (pH 7.4 at 25°C), 300 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.2 mg/mL BSA, 0.15% Triton X-100, and 50% (v/v) glycerol.|
|Storage Condition||-20 C|
|Recommended buffer for 100% activity||Optimal temperature||Enzyme activity in Thermo Scientific buffers, %||Tango buffer for double digestion|
1X / 2X
|EcoRI Buffer [Unique]||37°C||0-20||NR||100||100†||NR||100||2X|
† Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).
- Dam: never overlaps - no effect.
- Dcm: never overlaps - no effect.
- CpG: may overlap - cleavage impaired.
- EcoKI: never overlaps - no effect.
- EcoBI: may overlap - no effect.
|Methylation type||Sequence||Cleavage effect|
Cleavage efficiency close to the termini of PCR fragments
|bp from the recognition site to fragment end|
Number of recognition sites in DNA molecules
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|
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|
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