Recommended
buffer |
Reaction conditions |
|
Temperature |
Time |
| FastDigest® buffer |
37°C |
5 min |
Thermal Inactivation
FastDigest® EarI (Eam1104I) is inactivated by incubation at 80°C for
5 min.
Digestion of Plasmid DNA
1 µl of FastDigest® EarI (Eam1104I) 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® EarI (Eam1104I) 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® EarI (Eam1104I) 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® EarI (Eam1104I)
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
may overlap
never overlaps
may overlap |
- no effect.
- no effect.
- no effect.
- no effect.
- effect not determined. |
Note
Particular sites in lambda and plasmids DNA are difficult to cleave with
EarI (Eam1104I),
as well as with its prototype Ksp632I.
Recommendations for Double and Triple Digestions with FastDigest® Enzymes
Perform 5 minute double or triple digestions with FastDigest®
EarI (Eam1104I) 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.
