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Restriction Endonucleases >
Restriction Endonucleases >
AfeI |
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This product is currently
not available for online ordering.
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This native enzyme from SibEnzyme (V0213) has been discontinued and replaced by the NEB clone AfeI (R0652), Recombinant.
Recognition Site:
isoschizomers | compatible ends | single letter code
Source:
Alcaligenes faecalis T2774 (S.K. Degtyarev)
Reagents Supplied:
SEBuffer Y
Enzyme Properties
Activity in NEBuffers:
| NEBuffer 1: | | 25% | | NEBuffer 2: | | 50% | | NEBuffer 3: | | 25% | | NEBuffer 4: | | 100% |
When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
Methylation Sensitivity:
| dam methylation: Not sensitive | | dcm methylation: Not sensitive | | CpG methylation: Blocked |
Heat Inactivation:
65°C for 20 minutes
Survival in a Reaction:
Minimum units to digest 1 µg of substrate DNA in 16 hours: 0.25 unit(s)
Reaction & Storage Conditions
Reaction Conditions:
1X SEBuffer Y
Incubate at
37°C.
1X SEBuffer Y:
33 mM Tris-acetate
66 mM potassium acetate
10 mM Magnesium Acetate
1 mM Dithiothreitol
pH 7.9 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme required to digest 1 µg of λ (BamHI digest) DNA in 1 hour at 37°C in a total reaction volume of 50 µl.
Unit Assay Substrate:
λ DNA (BamHI digest)
Storage Conditions:
20 mM Tris-HCl
50 mM KCl
1 mM Dithiothreitol
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.5 @ 25°C
Storage Temperature:
-20°C
Diluent Compatibility:
Diluent A
Notes
General notes:- AfeI is an isoschizomer of Eco47III.
FAQs
- Why does AfeI's catalog number start with V instead of R?
- What buffers can be used for double digests?
- Does AfeI replave one previously sold?
- Are more units of AfeI required to cut supercoiled DNA than lambda DNA?
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Ligation and Re-cutting:
After a 10-fold overdigestion with AfeI, approximately 75%
of the DNA fragments can be ligated with T4 DNA Ligase (at a 5' termini concentration of 1-2 μM)
at 16ºC. Of these ligated fragments, approximately 75% can be recut with AfeI.
16-Hour Incubation:
A 50 μl reaction containing 1 μg of λDNA
and 20 units of AfeI incubated for 16 hours at 37ºC
resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.
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