Recommended
buffer |
Reaction conditions |
|
Temperature |
Time |
| FastDigest® buffer |
37°C |
5 min |
Thermal Inactivation
Phenol/chloroform extraction and ethanol precipitation of DNA. Thermal inactivation is not
applicable for FastDigest® SpeI (BcuI).
Digestion of Plasmid DNA
1 µl of FastDigest® SpeI (BcuI) digests up to 1 µg of plasmid DNA in 5 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® SpeI (BcuI) 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® SpeI (BcuI) digests 1 µg of genomic DNA in 5 min, or 5 µg of
genomic DNA in 30 min, see
Protocol for Fast Digestion of Genomic DNA with FastDigest® Enzymes.
Ligation
and Recleavage
After overdigestion with 1 µl of FastDigest® SpeI (BcuI) for 1 hour, more than 80% of the DNA fragments can be
ligated and more than 90% of these can be recut.
| Methylation
Effects |
Dam:
Dcm:
CpG:
EcoKI:
EcoBI: |
never overlaps
never overlaps
never overlaps
may overlap
may overlap |
|
- no effect.
- no effect.
- no effect.
- effect not determined.
- effect not determined. |
Compatible Ends
AvrII, Eco130I,
NheI, StyI, XbaI,
XmaJI.
Recommendations for Double and Triple Digestions with FastDigest® Enzymes
Perform 5 minute double or triple digestions with FastDigest®
SpeI (BcuI) 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.
