ScrFI

Description

Product Source

An E. coli strain that carries the cloned ScrFI gene from Streptococcus cremoris F (C. Daly)

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
CutSmart® Buffer-2010X

Properties and Usage

Unit Definition

One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Reaction Conditions

1X CutSmart® Buffer
Incubate at 37°C

1X CutSmart® Buffer:
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 μg/ml BSA
pH 7.9 @ 25°C

Activity in NEBuffers

NEBuffer 1.1: 100%
NEBuffer 2.1: 100%
NEBuffer 3.1: 100%
CutSmart® Buffer: 100%

Diluent Compatibility

Storage Temperature

-20°C

Storage Conditions

10 mM Tris-HCl
250 mM NaCl
0.1 mM EDTA
200 μg/ml BSA
50% Glycerol
1 mM DTT
0.15% Triton® X-100
pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 min

Methylation Sensitivity

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

Notes

  1. The recognition site for ScrFI is partially modified in most E. coli strains. Therefore in order to digest DNA grown in them (e.g. plasmids, phages), one must use a strain deficient in cytidine methylase.
  2. ScrFI produces DNA fragments that have a single-base 5´ extension which are more difficult to ligate than blunt-ended fragments.
  3. Star activity may result from extended digestion.

FAQs

  1. Do degenerate recognition sites need to be palindromic?
  2. My enzyme is no longer Time-Saver™ qualified. What happened?
  3. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
  4. How can I access the old NEBuffer™ Activity Chart?
  5. Why is my Restriction Enzyme not cutting DNA?
  6. Why do I see additional DNA bands on my gel after a restriction digest?
  7. Why do I see a DNA smear on an agarose gel after a restriction digest?
  8. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?

Tech Tips

This enzyme produces a single base extension, which is difficult to ligate.  This enzyme works in all NEB buffers.

Protocols

  1. Optimizing Restriction Endonuclease Reactions
  2. Double Digest Protocol with Standard Restriction Enzymes

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.
  • 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.