US Patent 7504240 Methods for synthesizing polynucleotides using a single primer

An objective of the present invention is to provide methods for synthesizing polynucleotides comprising an unknown nucleotide sequence in a region containing their 5′-end. By using a single gene-specific primer, nucleotide sequences complementary to each other are added respectively at the 5′-and 3′-ends of a desired polynucleotide, to synthesize a desired polynucleotide. Requisite nucleotide sequences can be easily added by incubating with a primer, double-stranded DNA, and template-dependent DNA polymerase. By applying the present invention to 5′ RACE, genes expressed in a low-level, which cannot be obtained by known methods, have been efficiently obtained.