Agarase

Detailní popis

Agarase specifically digests the agarose polysaccharide core into neoagaro-oligosaccharides (1). Agarase allows for gentle yet efficient recovery of DNA or RNA fragments from low melting point agarose. The recovered nucleic acids can be directly used for amplification, cloning, sequencing, etc.

Highlights

• Gentle – recovers intact DNA and RNA
• High recovery yields – > 90% of the starting DNA recovered following extraction
• Ideal for both long (> 30 kb) and short (< 100 bp) DNA fragments 
• Active in TAE and TBE electrophoresis buffers 

Applications

• Quantitative recovery of DNA or RNA from low melting point (LM) agarose gels.

Note

• Incubate at 42°C.
• Conventional agarose is not suitable for digestion by Agarase.
• Activity of agarase in different buffers (in comparison to activity in assay buffer (1x TBE)):
  • 1x TBE (90 mM Tris-borate, 2 mM EDTA, pH 8.3) - 100%.
  • 1x TAE (40 mM Tris-acetate, 1 mM EDTA, pH 8.5) - 120%.
  • 1x TPE (90 mM Tris-phosphate, 2 mM EDTA, pH 7.7) - 120%.
  • 1x Bis-Tris (50 mM Bis-Tris-HCl, 1 mM EDTA, pH 6.5) - 150%.

    CategoryName
    Definition of Activity Unit	One unit of the enzyme completely degrades 100 µL (approx. 100 mg) of molten 1% low melting point agarose in 30 min at 42°C. Enzyme activity is assayed in the 1x TBE buffer: 89 mM Tris base, 89 mM boric acid, 2 mM EDTA.
    Hazardous	No
    Hazardous:	No
    Inactivation	Inactivated by heating at 70°C for 10 min.
    Inhibition	Inhibitors: ethidium bromide at concentrations higher than 5 µg/mL, transition metal ions.
    Molecular Weight	32.7 kDa monomer.
    Quality Control	The absence of endo-, exodeoxyribonucleases, ribonucleases, and phosphatases confirmed by appropriate quality tests. Functionally tested in recovery of DNA fragments from low melting point agarose.
    Shelf Life:
    Shipping Condition:
    Shipping Information
    Source	E. coli cells with a plasmid containing the gene encoding beta-agarase from Pseudomonas atlantica.
    SpecificationName
    SpecificationValue
    Storage Buffer	The enzyme is supplied in 50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 0.1% (v/v) Triton X-100, and 50% (v/v) glycerol.
    Storage Condition	-20 C
    Storage Condition:	-20 C