EcoRV-HF® RE-Mix® is a ready to use 10X master mix combining the restriction enzyme, EcoRV-HF (NEB #R3195 ), NEBuffer, BSA and loading dye in a single tube. Simply add DNA and water and incubate for 15 minutes to perform the restriction digest.
Product SourceAn E. coli strain that carries the cloned and modified EcoRV gene from the plasmid J62 pLG74 (L.I. Glatman)
Reaction Definition1X EcoRV-HF RE-Mix and DNA in a total reaction volume of 20 μl. Incubate at 37°C for 15 minutes.
One reaction is defined as the amount of RE-Mix that will digest 1 μg of DNA in 15 minutes at 37°C in a total reaction volume of 20 μl.
Properties and Usage
Heat Inactivation65°C for 20 min
dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Impaired by Some Combinations of Overlapping
Relevant patent applications include: US2009-0029376, EP2390320, CN101802183A, IN 7681/CHENP/2009 and JP 2010-516300.
- What are the advantages of using a RE-Mix Restriction Enzyme Master Mix?
- For some restriction enzymes star activity can be a concern. Is this also applicable to the RE-Mix?
- The bands on my gel are not as sharp as I would like. Is there a way to improve the sharpness?
- Is it possible to use RE-Mix in double digests?
- Can RE-Mix be used in overnight digestions?
- Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
Usage Guidelines & Tips
- Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
- Activity of Restriction Enzymes in a Q5®, Taq or Phusion PCR Mix
- Cleavage Close to the End of DNA Fragments
- Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
- Double Digests
- Effects of CpG Methylation on Restriction Enzyme Cleavage
- Heat Inactivation
- Megabase Mapping
- NEBuffer Activity/Performance Chart with Restriction Enzymes
- Optimizing Restriction Endonuclease Reactions
- Optimizing Restriction Endonuclease Reactions When Using RE-Mix® Restriction Enzyme Master Mix
- Restriction Endonucleases - Survival in a Reaction
- Restriction Enzyme Diluent Buffer Compatibility
- Single Letter Codes
- Star Activity
- Traditional Cloning Quick Guide