β-Agarase I

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

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.

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.

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 Davis, T. and Guan, C. Unpublished observation

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

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 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.

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]

• M0392S_L_v1

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]

• M0392S_L_v1_0271508
• M0392S_L_v1_0271608
• M0392S_L_v1_0271701
• M0392S_L_v1_0271706
• M0392S_L_v1_0271801
• M0392S_L_v1_0281804
• M0392S_v1_10007348
• M0392S_v1_10030182
• M0392L_v1_10030184
• M0392S_v1_10033915
• M0392S_v1_10050911
• M0392L_v1_10050909
• M0392S_v1_10057643
• M0392L_v1_10057642
• M0392S_v1_10071611
• M0392L_v1_10071612
• M0392S_v1_10100140
• M0392L_v1_10100143
• M0392S_v1_10137954
• M0392L_v1_10137956
• M0392S_v1_10159995
• M0392L_v1_10176562
• M0392S_v1_10191395
• M0392L_v1_10205105
• M0392L_v1_10212041
• M0392L_v1_10236427
• M0392S_v1_10238105
• M0392S_v1_10273393
• M0392L_v1_10281714
• M0392L_v1_10310202
• M0392S_v1_10310216

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

• β-Agarase I
• β-Agarase I Reaction Buffer

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