Polynucleotides having in excess of 1,000 nucleotides can be prepared
using a solid phase synthesis technique. A feature of the technique is
the use of a reusable solid support that contains covalently bound
oligonucleotide. This covalently bound oligonucleotide is annealed to a
bridge oligonucleotide, where the bridge is also annealed to a first
oligonucleotide that forms a portion of the target polynucleotide. After
the target polynucleotide is synthesized, it can be removed from the
solid support under denaturing conditions, and the solid support re-used
to prepare additional target polynucleotides. The yield of the target
polynucleotide increases when shearing force is applied to the solid
support that is linked to the growing oligonucleotide. This shearing
force is thought to extend the growing end of the oligonucleotide away
from contact with other oligonucleotide bound to the solid support and
make that end more accessible to annealing with solution oligonucleotide.
The synthesis is conveniently accomplished on a porous frit, where
reagents and washing solutions are pumped through the frit.