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Agarase

LO certified Recombinant enzyme Thermal inactivation at 70°C in 10 min

Enzyme specifically digests the agarose polysaccharide core which allows for gentle yet efficient recovery of DNA or RNA fragments from low melting point agarose.
  
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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%.
     
  
Storage Condition-20 C
HazardousNo
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.
InactivationInactivated by heating at 70°C for 10 min.
InhibitionInhibitors: ethidium bromide at concentrations higher than 5 µg/mL, transition metal ions.
Molecular Weight32.7 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.
SourceE. coli cells with a plasmid containing the gene encoding beta-agarase from Pseudomonas atlantica.
Storage BufferThe 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.
Agarase-mediated recovery of DNA fragments from a 1% LM point agarose gel

Agarase-mediated recovery of DNA fragments from a 1% LM point agarose gel

Agarase-mediated recovery of DNA fragments from a 1% LM point agarose gel

Figure 1. Agarase-mediated recovery of DNA fragments from a 1% low melting point agarose gel.
Individual fragments of GeneRuler 1 kb DNA Ladder (lane M) were extracted from a 1% TopVision Low Melting Point Agarose gel. Sizes of DNA fragments (lanes 1-14): 10000, 8000, 6000, 5000, 4000, 3500, 3000, 2500, 2000, 1500, 1000, 750, 500, 250 bp.


DNA purification from low melting point agarose with Agarase

DNA purification from low melting point agarose with Agarase

DNA purification from low melting point agarose with Agarase

Figure 2. DNA purification from low melting point agarose with Agarase


References

  1. W. Yaphe, The use of agarase from Pseudomonas atlantica in the identification of agar in marine algae (Rhodophyceae). Can. J. Microbiol. 3(7), 987-993 (December 1957).

MSDS & Certificates

pdf   Agarase - MSDS
pdf   MSDS (JP) - Agarase