Product Class: Other

β-Agarase I
NEBU cloned at NEB recombinant 42 65 Heat

Product Introduction

  • For the extraction of DNA from agarose gels
  • Isolated from a recombinant source
  • DNase- and RNase-free
Catalog # Size Concentration
M0392S 100.0 units 1000 units/ml
M0392L 500.0 units 1000 units/ml

Product Information

Description

β-Agarase cleaves the agarose subunit, unsubstituted neoagarobiose [3,6-anhydro-a-L-galactopyranosyl-1-3-d-galactose] to neoagaro-oligosaccharides (1).

Product Source

Isolated from a strain of E. coli that carries a plasmid which encodes the β-Agarase I gene.
This product is related to the following categories:
Other Products,
DNA Cleanup and Gel Extraction Products,
DNA Gel Extraction Products,
This product can be used in the following applications:
Nucleic Acid Purification

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0392S     -20    
      β-Agarase I M0392SVIAL -20 1 x 0.1 ml 1,000 units/ml
      β-Agarase I Reaction Buffer B0392SVIAL -20 1 x 1.5 ml 10 X
  • M0392L     -20    
      β-Agarase I M0392LVIAL -20 1 x 0.5 ml 1,000 units/ml
      β-Agarase I Reaction Buffer B0392SVIAL -20 4 x 1.5 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to digest 200 μl of molten low melting point or NuSieve agarose to nonprecipitable neoagaro-oligosaccharides in 1 hour at 42°C.

Reaction Conditions

1X β-Agarase I Reaction Buffer
Incubate at 42°C

1X β-Agarase I Reaction Buffer
10 mM Bis-Tris-HCl
1 mM EDTA
(pH 6.5 @ 25°C)

Storage Buffer

50 mM Bis-Tris-HCl
1 mM EDTA
50% Glycerol
pH 6.5 @ 25°C

Heat Inactivation

65°C for 15 minutes

Application Features

  • β-Agarase I digests agarose, releasing trapped DNA and producing carbohydrate molecules which can no longer gel. β-Agarase I can be used to purify both large (> 50 kb) and small (< 50 kb) fragments of DNA from gels. The remaining carbohydrate molecules and β-Agarase I will not, in general, interfere with subsequent DNA manipulations such as restriction endonuclease digestion, ligation, and transformation.

Product Notes

  1. β-Agarase I retains activity for several hours at 40-45°C and is stabilized by the presence of agarose in the reaction.
  2. Only low melting point agarose is suitable forβ-Agarase I digestion as the solution must be liquid at the incubationtemperature of 42°C. If the temperature falls below 42°C during the reactiontime, even low melting point agarose will begin to congeal and beundigestable.
  3. β-Agarase I is quickly inactivated at temperaturesabove 45°C. Therefore, when working with large volumes, be sure to leave ampletime for the molten agar to equilibrate to 42°C.
  4. β-Agarase I works best on gels made with Tris-acetatebuffer (TAE). For gels made with Tris-borate buffer (TBE), doubling the requiredamount of β-Agarase I is recommended.
  5. β-Agarase I works most efficiently on solutionscontaining 1% agarose or lower. For maximum digestion of higher percentage gels,melt the gel slice at 65°C and adjust the volume with 1X β-Agarase I Buffer sothat the final concentration of agarose is 1%.
  6. β-Agarase I exhibits optimal activity at pH 6.5.Greater than 75% of the optimal activity is maintained between pH5.0-8.5.
  7. Incubation at 95°C for 2 minutes or incubation at 65°C for 15 minutes inactivates 50 units of β-Agarase I. β-Agarase I retains activity for several hours at 40-45°C and is stabilized by the presence of agarose in the reaction.

References

  1. Yaphe, W. (1957). Can. J. Microbiol. 3, 987-993.
  2. Davis, T. and Guan, C. Unpublished observation

Protocols, Manuals & Usage

Protocols

  1. DNA Purification from Agarose Gels using Beta Agarase I (NEB #M0392)

FAQs & Troubleshooting

FAQs

  1. Can ligation and other DNA manipulations be carried out on β-Agarase I treated gel slices containing DNA?
  2. What is the molecular weight of β-Agarase I?
  3. What type of agarose will β-Agarase I digest?
  4. Will β-Agarase I work in other buffers?
  5. Why does a white precipitate form after the reaction using β-Agarase I?
  6. Can a high percentage low melt gel be digested with β-Agarase I?
  7. What is a common cause of β-Agarase I reaction failure?
  8. Does the gel running buffer have an effect on the β-Agarase I reaction?
  9. How can β-Agarase I be heat inactivated?
  10. How stable is β-Agarase I in reaction?
  11. What is the optimal pH for β-Agarase I digestion?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

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]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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