Inserting a tag sequence into a nucleic acid comprises using an anchor oligonucleotide comprising a hybridizing anchor sequence and a nonhybridizing tag-template sequence

摘要

Inserting a tag sequence into a nucleic acid comprises hybridizing an anchor sequence of an anchor oligonucleotide to a sequence segment of a template nucleic acid and synthesizing a new nucleic acid strand that is partially complementary to the template and has a 3' terminal sequence complementary to the nonhybridizing part of the anchor oligonucleotide, thereby forming a double-stranded region. Independent claims are also included for: (1) nucleic acid that is synthesized in a first-strand synthesis and has a 3' terminal tag sequence introduced in a matrix independent manner during first-strand synthesis, where the sequence complementary to the tag sequence is not part of a template nucleic acid; (2) anchor oligonucleotide having an anchor sequence in the 3' region and a tag-template sequence in the 5' region such that the anchor sequence is hybridized and the tag-template sequence is not hybridized to a template nucleic acid during second-strand synthesis, where synthesis of the second strand, which is complementary to part of the template, starts 3' to the hybridization site of the anchor sequence on the template strand and ends at the 5' end of the tag-template sequence by means of a polymerase template switch.