{"product_id":"neb-m0392l","title":"New England Biolabs, M0392L, β-Agarase I","description":"\u003cb\u003eRelated Categories\u003c\/b\u003e Other,, DNA Gel Extraction  \u003cb\u003eApplications\u003c\/b\u003e Nucleic Acid Purification  \u003cb\u003eSpecification\u003c\/b\u003e \u003cb\u003eUnit Definition\u003c\/b\u003e 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.  \u003cb\u003eReaction Conditions\u003c\/b\u003e 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)  \u003cb\u003eStorage Buffer\u003c\/b\u003e 50 mM Bis-Tris-HCl 1 mM EDTA 50% Glycerol pH 6.5 @ 25°C  \u003cb\u003eHeat Inactivation\u003c\/b\u003e 65°C for 15 minutes  \u003cb\u003eFAQ\u003c\/b\u003e Q: Can ligation and other DNA manipulations be carried out on β-Agarase I treated gel slices containing DNA? A: Yes. Q: What is the molecular weight of β-Agarase I? A: 30,000 Daltons Q: What type of agarose will β-Agarase I digest? A: Only low melting point agarose is suitable for β-Agarase I digestion as the solution must be liquid at the incubation temperature of 42°C. If the temperature falls below 42°C during the reaction time, even low melting point agarose will begin to congeal and be undigestable. Q: Will β-Agarase I work in other buffers? A: Yes. β-Agarase I will work in water and other buffers. The β-Agarase I concentration should be doubled for full activity in other buffers. The EDTA in the β-Agarase I buffer protects the DNA from a low level of nicking seen in buffers containing magnesium. Q: Why does a white precipitate form after the reaction using β-Agarase I? A: The reaction did not go to completion. See the questions below to optimize the reaction. Q: Can a high percentage low melt gel be digested with β-Agarase I? A: β-Agarase I works most efficiently on solutions containing 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 so that the final concentration of agarose is 1%. Q: What is a common cause of β-Agarase I reaction failure? A: Working at the wrong temperature. β-Agarase I is quickly inactivated at temperatures above 45°C. The low melting point agarose must be melted at 65°C then equilibrated at 42°C. Therefore, when working with large volumes, be sure to leave ample time for molten agar to equilibrate to 42°C. At lower temperatures the agarose starts to solidify making it resistant to digestion. Q: Does the gel running buffer have an effect on the β-Agarase I reaction? A: β-Agarase I works best on gels made with Tris-acetate buffer(TAE). For gels made with Tris-borate buffer (TBE), doubling the required amount of β-Agarase I is recommended. Q: How can β-Agarase I be heat inactivated? A: Incubation at 95°C for 2 minutes or incubation at 65°C for 15 minutes inactivates 50 units of β-Agarase I. Q: How stable is β-Agarase I in reaction? A: β-Agarase I retains activity for several hours at 45-50°C and is stabilized by the presence of agarose in the reaction. Q: What is the optimal pH for β-Agarase I digestion? A: β-Agarase I exhibits optimal activity at pH 6.5. Greater than 75% of the optimal activity is maintained between pH 5.0 - 8.5. ","brand":"New England Biolabs","offers":[{"title":"Default Title","offer_id":46835502055593,"sku":"M0392L","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-m0392l","provider":"Iright","version":"1.0","type":"link"}