KpnI-HF® RE-Mix® is a ready to use 10X master mix combining the restriction enzyme, KpnI-HF (NEB #R3142 ), 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 KpnI gene from Klebsiella pneumoniae OK8 (ATCC 49790)
Reaction Definition1X KpnI-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
dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive
Quality Control AssaysThe following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
- Endonuclease Activity (Nicking):
The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
- Exonuclease Activity (Radioactivity Release):
The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
- Ligation and Recutting (Terminal Integrity):
After an over-digestion of DNA with a restriction endonuclease the percentage of the DNA fragments ligated with T4 DNA ligase and the percentage that can be recut are determined by agarose gel electrophoresis.
- Non-Specific DNase Activity (16 hour):
The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.
US 2014/0154676, EP 2395079 (FR, GB, LT, DE 602008036313.9)
- 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?