how to extract restriction enzymes for bateria
This enzyme can be extracted and purified by using known techniques commonly employed for restriction endonucleases. The collected microbial cells are dispersed in a suitable buffer, and then broken down by ultrasonic treatment to allow extraction of the endonuclease by the buffer solution. After removal of the residue by ultracentrifugation, ammonium sulfate was added to the supernatant for salting out, and the precipitate which separated out was dissolved in a potassium phosphate buffer ( pH: 7.5 ) and dialyzed against a buffer of the same composition. The dialyzate was purified by ion-exchange chromatography on phosphocellulose and DEAE-cellulose, and by affinity chromatography on heparin-Sepharose, giving the endonuclease of this invention.
vinegar and sodium acetate are commonly availible chemicals that can be mixed to produce a buffer.
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