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Restriction Endonucleases >
Restriction Endonucleases >
BaeI |
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Prices are in US dollars and valid only for US orders.
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Recognition Site:
isoschizomers | compatible ends | single letter code
Source:
Bacillus sphaericus (H. Kong)
Reagents Supplied:
NEBuffer 2 (10X)
BSA (100X)
S-adenosylmethionine (SAM) (32 mM)
Enzyme Properties
Activity in NEBuffers:
| NEBuffer 1: | | 50% | | NEBuffer 2: | | 100% | | NEBuffer 3: | | 50% | | NEBuffer 4: | | 75% |
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 by some combinations of overlapping |
Activity at 37°C:
20%
Heat Inactivation:
65°C for 20 minutes
Survival in a Reaction:
Minimum units to digest 1 µg of substrate DNA in 16 hours: 0.50 unit(s)
Reaction & Storage Conditions
Reaction Conditions:
1X NEBuffer 2
Supplemented with 100 μg/ml Bovine Serum Albumin and 20 μM S-adenosylmethionine (SAM)
Incubate at
25°C.
1X NEBuffer 2:
10 mM Tris-HCl
50 mM NaCl
10 mM MgCl2
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 Adenovirus-2 DNA in 1 hour at 25°C in a total reaction volume of 50 µl.
Concentration:
5,000 units/ml
Unit Assay Substrate:
Adenovirus-2 DNA
Storage Conditions:
10 mM Tris-HCl
50 mM KCl
1 mM Dithiothreitol
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C
Storage Temperature:
-20°C
Diluent Compatibility:
Diluent A
Notes
General notes:- BaeI cleaves DNA substrates twice to excise its recognition site generating a 28 base-pair fragment with 5 base 3´ overhangs.
- In one sequence context, BaeI was found to cut at its defined cleavage site as well as one base pair further from its recognition site.
- Requires S-adenosylmethionine for optimal activity (supplied with enzyme).
FAQs
- Does BaeI have any special requirements?
- Does BaeI cleave DNA twice?
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
16-Hour Incubation:
A 50 μl reaction containing 1 μg of DNA
and 8 units of BaeI incubated for 16 hours at 25ºC
resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.
Exonuclease Activity:
Incubation of a 50 μl reaction containing 8 units of BaeI with 1 μg of a mixture of single and double-stranded
[3H] E. coli DNA (205 cpm/μg) for 4 hours at 25ºC
released < 0.01% of the total radioactivity.
Reagents Sold Separately
NEBuffer 2
BSA
S-adenosylmethionine (SAM)
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