NcoI-HF® RE-Mix®

Description

  
NcoI-HF® RE-Mix® is a ready to use 10X master mix combining the restriction enzyme, NcoI-HF (NEB #R3193 ), 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 Source

An E. coli strain that carries the cloned and modified NcoI gene from Nocardia corallina (ATCC 19070)

Reaction Definition

1X NcoI-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 Inactivation

80°C for 20 min

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive

FAQs

  1. What are the advantages of using a RE-Mix Restriction Enzyme Master Mix?
  2. For some restriction enzymes star activity can be a concern. Is this also applicable to the RE-Mix?
  3. The bands on my gel are not as sharp as I would like. Is there a way to improve the sharpness?
  4. Is it possible to use RE-Mix in double digests?
  5. Can RE-Mix be used in overnight digestions?
  6. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?

Protocols

  1. Standard Digest Using RE-Mix®
  2. Double Digest Protocol using Two RE-Mix® Enzymes
  3. Double Digest Protocol using One RE-Mix and One Standard Restriction Enzyme

Selection Charts

Usage Guidelines & Tips

Troubleshooting Guides

Interactive Tools

Quality Control

Quality Control Assays

The 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. Further information regarding NEB product quality can be found here.
  • 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.

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.