Product Class: Restriction Enzyme Re-Mix

AgeI-HF® RE-Mix®

re-mix icon cloned at neb recombinant engineered timesaver 5min incubation temp heat inactivation cpg
R5552_v1_000014
Catalog #SizeConcentration
R5552S25 reactions10X

Description

  
AgeI-HF® RE-Mix® is a ready to use 10X master mix combining the restriction enzyme, AgeI-HF® (NEB #R3552), NEBuffer, 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 AgeI gene from Ruegeria gelatinovora (ATCC 25655)

Reaction Definition

1X AgeI-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

65°C for 20 min

Methylation Sensitivity

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

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.
  • 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.

Supporting Documents

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
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.
  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?
  1. Standard Digest Using RE-Mix®
  2. Double Digest Using RE-Mix®