Insertion of sequence elements into nucleic acids

摘要

The present invention concerns a method for inserting one or more tag sequences into a nucleic acid characterized by the following steps: (a) preparation of a template nucleic acid; (b) hybridization of at least one anchor sequence of at least one anchor oligonucleotide with one sequence section of the template nucleic acid; and (c) synthesization of a new strand of nucleic acid, which is partially complementary to the template nucleic acid and which contains a sequence complementary to the non-hybridized portion of the anchor oligonucleotide, e.g. to at least one tag sequence, on its 3′ end.

主权项

1. A method for random insertion of a tag sequence into a nucleic acid comprising:
in a first strand synthesis reaction, (a) providing a template nucleic acid and a primer, (b) hybridizing an anchor oligonucleotide to the template nucleic acid,
wherein the anchor oligonucleotide comprises an anchor sequence and a template tag sequence,wherein the anchor sequence is a random sequence,andwherein the template tag sequence is a non-hybridized portion of the anchor oligonucleotide; and (c) extending the primer of step (a), along the template nucleic acid to form a new strand of nucleic acid,
wherein a portion of the new strand of nucleic acid comprises a sequence complementary to the template nucleic acid, andwherein a portion of the resulting new strand of nucleic acid comprises a sequence complementary to the template tag sequence of the anchor oligonucleotidethereby randomly inserting a sequence complementary to the template tag sequence of the anchor oligonucleotide into a new strand of nucleic acid.