Product Class: Restriction Endonuclease

BaeI

cloned at neb recombinant timesaver 5min incubation temp heat inactivation sam cpg
Bae-I-cutsite_1
Catalog #SizeConcentration
R0613S250 units5,000 units/ml
R0613L1,250 units5,000 units/ml

Description

Product Source

An E. coli strain that carries the cloned BaeI gene from Bacillus sphaericus SDZ-8 (R. Morgan)

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
S-adenosylmethionine (SAM)-2032 mM
CutSmart® Buffer-2010X

Properties and Usage

Unit Definition

One unit is defined as the amount of enzyme required to digest 1 µg of Lambda DNA in 1 hour at 25°C in a total reaction volume of 50 µl.

Reaction Conditions

1X CutSmart® Buffer
Supplement with 20 μM S-adenosylmethionine (SAM)
Incubate at 25°C

1X CutSmart® Buffer:
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 μg/ml BSA
pH 7.9 @ 25°C

Activity in NEBuffers

NEBuffer 1.1: 50%
NEBuffer 2.1: 100%
NEBuffer 3.1: 50%
CutSmart® Buffer: 100%

Diluent Compatibility

Storage Temperature

-20°C

Storage Conditions

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
200 μg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 min

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Blocked by Some Combinations of Overlapping

Heat Inactivated

Yes

Quality Control

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
  • Exonuclease Activity (Radioactivity Release):
    The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
  • Ligation and Recutting (Terminal Integrity):
    After an over-digestion of DNA with a restriction endonuclease the percentage of the DNA fragments ligated with T4 DNA ligase and the percentage that can be recut are determined by agarose gel electrophoresis.

Supporting Documents

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Notes

  1. BaeI cleaves DNA substrates twice to excise its recognition site generating a 28 base-pair fragment with 5 base 3´ overhangs.
  2. BaeI was found to cut at its defined cleavage site as well as one base pair further from its recognition site.
  3. Requires S-adenosylmethionine for optimal activity (supplied with enzyme).
  1. Does BaeI have any special requirements?
  2. Does BaeI cleave DNA twice?
  3. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
  4. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  5. How can I access the old NEBuffer Activity Chart?
  6. I tested your restriction enzyme on the substrate DNA recommended by NEB, and it appears to be active, however it does not digest my DNA. What could be the reason?
  7. Why is my Restriction Enzyme not cutting DNA?
  1. Optimizing Restriction Endonuclease Reactions
  2. Double Digest Protocol with Standard Restriction Enzymes
  3. Time-Saver Protocol for Restriction Enzyme Digests