Product SourceAn E. coli strain that carries the cloned FatI gene from Flavobacterium aquatile NL3 (S.K. Degtyarev)
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
Properties and Usage
Unit DefinitionOne 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.
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 NEBuffersNEBuffer 1.1: 10%
NEBuffer 2.1: 100%
NEBuffer 3.1: 50%
CutSmart™ Buffer: 50%
10 mM Tris-HCl
50 mM NaCl
0.1 mM EDTA
1 mM DTT
200 μg/ml BSA
pH 7.4 @ 25°C
Heat Inactivation80°C for 20 min
dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive
Quality Control AssaysThe 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.
- 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.
- FatI is a neoschizomer of NlaIII.
- 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?
- Does FatI have any neoschizomers?
- What is the activity of FatI at 37°C?
- What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
- How can I access the old NEBuffer Activity Chart?
- How can I access the old Double Digest Finder?
- What effect does BSA have on the performance of NEB's restriction enzymes when included in the new buffers?