Recommended
buffer |
Reaction conditions |
|
Temperature |
Time |
| FastDigest® buffer |
37°C |
5 min |
Thermal Inactivation
FastDigest® NotI is inactivated by incubation at 80°C for 5 min.
Digestion of Plasmid DNA
1 µl of FastDigest® NotI digests up to 0.4 µg of plasmid DNA in 5 min or
1 µg in 30 min, see
Protocol for Fast
Digestion and Analysis of Plasmid DNA with FastDigest® Enzymes
and
Protocol for Fast Vector
Preparation with FastDigest® Enzymes.
Digestion of PCR Products
1 µl of FastDigest® NotI digests ~0.2 µg of PCR product in 5 min,
see
Protocol for Fast Digestion of PCR
Products with FastDigest® Enzymes.
Digestion of Genomic DNA
1 µl of FastDigest® NotI digests 1 µg of genomic DNA in 10 min, or 5 µg of
genomic DNA in 60 min, see
Protocol for Fast Digestion of Genomic DNA with FastDigest® Enzymes.
Ligation
and Recleavage
After overdigestion with 1 µl of FastDigest® NotI for 1 hour, more than 95% of DNA fragments can
be ligated and recut.
| Methylation
Effects |
Dam:
Dcm:
CpG:
EcoKI:
EcoBI: |
never overlaps
never overlaps
completely overlaps
never overlaps
never overlaps |
|
- no effect.
- no effect.
- blocked.
- no effect.
- no effect. |
Compatible Ends
Bsp120I, CfrI,
Eco52I.
Recommendations for Double and Triple Digestions with FastDigest® Enzymes
Perform 5 minute double or triple digestions with FastDigest® NotI and any other FastDigest® enzyme(s) in FastDigest® buffer,
see Protocol for
Double Digestion with FastDigest® Enzymes and
Protocol for Triple Digestion with
FastDigest® Enzymes.
