| 摘要 |
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.
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