Product Class: Restriction Endonuclease


icon_cloned icon_recomb icon_neb2_1 dil_A-grey icon_incTemp55 heat inactivation

Product Introduction

Having supplied restriction enzymes to the research community for over 40 years, NEB has earned the reputation of being the leader in enzyme technologies.  Working continuously to be worth of that distinction, NEB strives to develop enzyme of the highest purity and unparalleled quality. 

Catalog # Size Concentration
R0650L 250 units 2,000 units/ml
R0650S 50 units 2,000 units/ml

Product Information


Product Source

An E. coli strain that carries the cloned FatI gene from Flavobacterium aquatile NL3 (S.K. Degtyarev)

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration
NEBuffer™ 2.1 -20 10 X
Product Categories:
Restriction Endonucleases: C-G
Restriction Enzyme Digestion

Properties and Usage

Unit Definition

One unit is defined as the amount of enzyme required to digest 1 µg of pUC19 DNA in 1 hour at 55°C in a total reaction volume of 50 µl.

Reaction Conditions

1X NEBuffer™ 2.1
Incubate at 55°C

1X NEBuffer™ 2.1
50 mM NaCl
10 mM Tris-HCl
10 mM MgCl2
100 μg/ml BSA
(pH 7.9 @ 25°C)

Activity in NEBuffers

CutSmart® Buffer: 50%
NEBuffer 1.1: 10%
NEBuffer 2.1: 100%
NEBuffer 3.1: 50%

Diluent Compatibility

Storage Temperature


Storage Conditions

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

Heat Inactivation

80°C for 20 min

Methylation Sensitivity

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

Activity at Temperature

@37°C: 20%


  1. FatI is a neoschizomer of NlaIII.

Faqs & Tech Tips


  1. How many base pairs should be added at the end of a PCR primer after the FatI recognition site to guarantee that FatI will cut properly?
  2. Does FatI have any neoschizomers?
  3. What is the activity of FatI at 37°C?
  4. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
  5. How can I access the old NEBuffer Activity Chart?
  6. What effect does BSA have on the performance of NEB's restriction enzymes when included in the new buffers?
  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 & Manuals


  1. Optimizing Restriction Endonuclease Reactions
  2. Double Digest Protocol with Standard Restriction Enzymes
  3. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
  4. Protocol for Digestion Prior to droplet digital PCR (ddPCR)

Other Tools & Resources

Selection Charts

Usage Guildelines & Tips

Troubleshooting Guides

Restriction Enzyme Troubleshooting Guide


Quality & Safety

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.


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]

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]

Safety DataSheets

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