Polynucleotide Celluloses as Solid State Primers and Templates for Polymerases

An oligomer of deoxyribothymidylate in 5'-ester linkage to cellulose particles serves as a primer and template for the Escherichia coli DNA polymerase, as a template for RNA polymerase, and as an initiator for the calf thymus terminal deoxynucleotidyltransferase. Synthesis by DNA polymerase of the homopolymer pair, polydeoxyriboadenylate and polydeoxyribothymidylate, takes place when oligodeoxyribothymidylate cellulose is hydrogen-bonded to an oligodeoxyriboadenylate to form a template pair. The product contains deoxythymidylate covalently linked to the cellulose and deoxyadenylate hydrogen-bonded to the deoxythymidylate. Block copolymers formed by transferase extension of an oligonucleotide cellulose can form hybrids with other synthetic polymers according to base pair specificities. Single stranded DNA extended with a homopolymer by terminal transferase can hydrogen bond via the homopolymer region to a complementary derivative of polynucleotide cellulose. Upon replication with DNA polymerase, the product strand is covalently attached to the cellulose. The advantages of the polynucleotide cellulose system include the known polarity of primer and product, the ease of quantitative collection and separation of the insoluble template and complementary product, and the capacity to distinguish between reactions occurring free in solution and those involving components fixed to a solid state matrix.