ABSTRACT
Linkage of polynucleotides to insoluble or soluble polymer supports offers many advantages in studies of the enzymic reactions. DNA polymerase, RNA polymerase, and terminal nucleotidyltransferase can interact with polynucleotides linked to 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 the solution and those involving components fixed to a solid-state matrix. In a primer-dependent replication, the reaction stops when the duplex has been completed and further synthesis needs separation of the duplex and a restart in the presence of an excess of the primer. Ovalbumin mRNA was purified from hen oviduct polysomal RNA by poly(U)-Sepharose chromatography. The ovalbumin mRNA comprised about 60–65% of this total messenger preparation. DNA complementary to ovalbumin mRNA and covalently bound to cellulose has been synthesized.
