| Products | beta-Agarase from USB Corp. | |
| Company | USB Corp. | |
| Item | beta-Agarase | |
| Price | ||
| Description | Source: Pseudomonas atlantica Description: Beta-Agarase is useful in recovering DNA and RNA fragments from low-melting-temperature agarose. Beta-Agarase digests the agarose into oligo- and disaccharides. Tested User Friendly Properties: Heat Inactivation: 65C, 15 min Molecular Weight: 30,000 Da Optimum pH: 6.5 Purity: Tested for contaminating endonucleases, exonucleases and ribonucleases. Storage Buffer: 50mM Bis-Tris-HCl (pH 6.5), 1mM EDTA, 50% glycerol. Shipping and Storage: Shipped on dry ice. Store at -20C. Assay Conditions: Reaction mixture contains 10mM Bis-Tris-HCI (pH 6.5) and 1mM EDTA. Unit Definition: One unit is the amount of enzyme required to digest 200 l of melted low-melting-point agarose to oligo- and disaccharides in 1 hr at 40C. Concentration: 1 unit/l Tested User FriendlyTM Functional Test: Isolation of linearized plasmid from low-melting agarose, > 50% recovery. Applications: * Quantitative recovery of DNA from low-melting point agarose gels. PROTOCOL FOR RECOVERING FRAGMENTS FROM LOW-MELT AGAROSE *Equilibrate DNA/agarose in at least 2 volumes of 1X Beta-Agarase Reaction Buffer for approximately 20 min. *Spin briefly and remove the supernatant. *Incubate DNA/agarose at 70C for 10 min to melt the agarose. *Cool to 40C and maintain the temperature at 37C. *Add an equal volume of 1X Beta-Agarase Reaction Buffer. *Add one unit of Beta-Agarase for every 50 l of agarose gel. Do not include the volume of the buffer for this determination. *Incubate the enzyme/DNA/agarose for 1-18 hours at 37C. The treated DNA may be used directly for ligation, restriction digests, etc. or further purified as in steps 8-12. *Extract once with buffer equilibrated phenol. *Extract once with buffer equilibrated phenol:chloroform (1:1). *Extract twice with chloroform:isoamyl alcohol (24:1). *Add 2 volumes of TE (pH 7.6) to prevent precipitation of the oligosaccharides. Precipitate with 0.05 volumes of NaCl and 2 volumes of 95% ethanol. Incubate at -20C for 30 min, then spin in a microcentrifuge. Wash the pellet twice with 70% ethanol. *The DNA solution may be used directly for ligation, restriction digests, transformation and sequencing or the purified DNA may be concentrated. References: *YAPHE, W. (1957) Can. J. Microbiol. 3, 987-993. *BURMEISTER, M. AND LEHRACH, H. (1986) Nature 324, 582-585. Functionally Tested 10X Beta-Agarase Reaction Buffer (1 ml included, PN 73432): 100mM Bis-Tris-HCl (pH 6.5), 10mM EDTA | |
| Info | USB Corp.26111 Miles Road Cleveland Ohio 44128
Fax Number: 216.464.5075 Web Site: http://www.usbweb.com |