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NEBuffer 2
Bme1580I
NEBuffer 237Heat Inactivated
Nomenclature Update
This product is currently not available for online ordering.
This product has been discontinued and replaced by BaeGI (R0708)
Download:MSDS PDF


Recognition Site:

GKGCMC

isoschizomers | compatible ends | single letter code

Source:
Bacillus megaterium 1580 (C. Nkenfou)

Reagents Supplied:
NEBuffer 2


Enzyme Properties


Activity in NEBuffers:
NEBuffer 1:75%
NEBuffer 2:100%
NEBuffer 3:75%
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: Not sensitive

Heat Inactivation:
80°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
Incubate at 37°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 λ DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Concentration:
5,000 units/ml

Unit Assay Substrate:
λ 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:
  1. Bme1580I is an isoschizomer of BseSI.

FAQs


  1. Do degenerate recognition sites need to be palindromic?

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 5-fold overdigestion with Bme1580I, > 95% 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, > 95% can be recut with Bme1580I.


16-Hour Incubation:
A 50 μl reaction containing 1 μg of λDNA and 25 units of Bme1580I incubated for 16 hours at 37ºC 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 50 units of Bme1580I with 1 μg of a mixture of single and double-stranded [3H] E. coli DNA (105 cpm/μg) for 4 hours at 37ºC released < 1% of the total radioactivity.


Reagents Sold Separately


NEBuffer 2

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