Patent ID: 6175006
Filing Date: 2001-01-16
Classification: B01J,C07H,C12N,C40B

Abstract:
A method for purifying a polynucleotide comprising(a) forming a polynucleotide on a solid support by repeated steps of (i) deblocking a 5'-hydroxyl group of a 5'-blocked protected nucleoside having a 3'-hydroxyl and a 5'-hydroxyl, (ii) reacting the deblocked 5'-hydroxyl with a 5'-blocked protected nucleoside monomer selected from the group consisting of 5'-blocked protected nucleoside-3'-phosphoramidite and 5'-blocked protected nucleoside-3'-hydrogen phosphonate to form either a protected correct-sequence polynucleotide chain or a failure sequence, the failure sequence having a 5'-hydroxyl, and (iii) repeating steps (i) and (ii) until a desired polynucleotide is obtained, wherein the solid support comprises a chamber containing (1) a nonswellable porous polystyrene support capable of remaining substantially mechanically rigid during polynucleotide synthesis, and (2) a hydrophobic adsorbent capable of non-covalently adsorbing a tritylated polynucleotide,(b) cleaving the polynucleotide from the nonswellable porous polystyrene support to form a cleavage mixture,(c) purifying the cleaved polynucleotide from the cleavage mixture by adsorbing the cleaved polynucleotide to the hydrophobic adsorbent and washing the hydrophobic adsorbent to selectively remove unblocked polynucleotides which the cleaved polynucleotide, and(d) deblocking the 5'-hydroxyl of the adsorbed polynucleotide to form a deblocked polynucleotide and eluting the deblocked polynucleotide from the hydrophobic adsorbent.