NheI-HF® RE-Mix®

This product has been discontinued on July 28, 2016. R3131 is a suitable alternative for most applications.
  • This product was discontinued on 07/28/2016
re-mix icon cloned at neb recombinant engineered timesaver 5min incubation temp heat inactivation cpg
R5131_v1_000032
Isoschizomers
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Discontinued Products

Description

  
NheI-HF® RE-Mix® is a ready to use 10X master mix combining the restriction enzyme, NheI-HF (NEB #R3131 ), 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 NheI gene from Neisseria mucosa heidelbergensis (ATCC 25999).

Reaction Definition

1X NheI-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

Storage Temperature

-20°C

Heat Inactivation

80°C for 20 min

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Blocked by Some Combinations of Overlapping

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?
  7. Why is my Restriction Enzyme not cutting DNA?
  8. Why do I see a DNA smear on an agarose gel after a restriction digest?
  9. Why do I see additional DNA bands on my gel after a restriction digest?
  10. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?

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
  4. Time-Saver Protocol for Restriction Enzyme Digests

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

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.

Certificate of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Safety Data Sheet

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

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.