| Recognition site | YA↑TR
RT↓AY |
| Source | Flavobacterium aquatile B15 |
| Assayed on | Double-stranded oligonucleotide
5`- CGAGTTCA^TAGCTGGGCCCAAC -3`
3`- GCTCAAGT^ATCGACCCGGGTTG -5` |
| Description | Note! FaiI cleaves 4 expected recognition sites as well as several other sites with a weaker activity.
In the case of long incubation with FaiI DNA can be digested to small oligos. |
| Unit definition | One unit is defined as the amount of enzyme required to cleave 1 pmol of the double-stranded oligonucleotide of the above indicated structure in 1 hour at 50°C in a total reaction volume of
20 μl. |
| Optimal SE-buffer | B (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 1 mM DTT.) |
| Enzyme activity (%) | | B | G | O | W | Y | Rose | | 100 | 50 - 75 | 10 - 25 | 25 - 50 | 25 - 50 | 100 |
|
| Optimal temperature | 50oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 100 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C. |
| Ligation | After 3-fold overdigestion with enzyme about 90% of the pUC19 DNA fragments can be ligated with DNA Ligase and recut. |
| Reagents Supplied with Enzyme | 10 X SE-buffer B |
| Inactivation 20 minutes under | 80oC |
| Quality control | Restriction Endonucleases: Quality Control |
NGS Application:
FaiI may be used for DNA fragmentation in NGS |
Hydrolysis in 5% polyacrylamide gele with TAE buffer
Lanes:
1. DNA HeLa + 1μ FaiI;
2. DNA HeLa + 1/2μ FaiI;
3. DNA HeLa + 1/4μ FaiI;
4. DNA HeLa (control);
5. SE 1 Kb DNA Ladder;
6. DNA HeLa + 8μ GlaI;
7. DNA HeLa + 8μ GlaI + 1μ FaiI;
8. DNA HeLa + 8μ GlaI + 1/2μ FaiI;
9. DNA HeLa + 8μ GlaI + 1/4μ FaiI;
10. SE 1 Kb DNA Ladder;
11. pUC19/MspI Ladder.
Hydrolysis in 1.2% agarose gele with TAE buffer
Lanes:
1. DNA HeLa + 1μ FaiI;
2. DNA HeLa + 1/2μ FaiI;
3. DNA HeLa + 1/4μ FaiI;
4. DNA HeLa (control);
5. SE 1 Kb DNA Ladder;
6. DNA HeLa + 8μ GlaI;
7. DNA HeLa + 8μ GlaI + 1μ FaiI;
8. DNA HeLa + 8μ GlaI + 1/2μ FaiI;
9. DNA HeLa + 8μ GlaI + 1/4μ FaiI.
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| MSDS: | Download MSDS as PDF |
| References: |  Tarasova G.V., Chernukhin V.A., Tomilova J.E., Degtyarev S.Kh Substrate specificity of new restriction endonuclease FaiI // In press
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