Compositions including a double stranded nucleic acid molecule and a stem-loop oligonucleotide
AssigneeTakara Bio USA, Inc.
InventorsVladimir L. Makarov, Emmanuel Kamberov, Brendan J. Tarrier
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Abstract
The present invention concerns preparation of DNA molecules, such as a library, using a stem-loop oligonucleotide. In particular embodiments, the invention employs a single reaction mixture and conditions. In particular, at least part of the inverted palindrome is removed during the preparation of the molecules to facilitate amplification of the molecules. Thus, in specific embodiments, the DNA molecules are suitable for amplification and are not hindered by the presence of the palindrome.
Claims20
An oligonucleotide-attached nucleic acid molecule comprising:
- a double stranded nucleic acid molecule comprising a first strand and a second strand;
- and
- a stem-loop oligonucleotide comprising:
- an inverted repeat;
- a loop;
- a non-replicable base or sequence, or at least one deoxy-uridine;
- and
- a 5′ end that lacks a phosphate;
- wherein the stem-loop oligonucleotide is attached to the first strand of the double stranded nucleic acid molecule and the 5′ end of the stem-loop oligonucleotide is separated from the second strand of the double stranded nucleic acid molecule.
An oligonucleotide-attached nucleic acid molecule comprising:
- a double stranded nucleic acid molecule and a stem-loop oligonucleotide comprising an inverted repeat and a loop, wherein one strand of the stem-loop oligonucleotide is attached to one strand of the double stranded nucleic acid molecule and the stem loop oligonucleotide further comprises at least one deoxy-uridine.
A composition comprising a plurality of oligonucleotide-attached nucleic acid molecules each comprising:
- a double stranded nucleic acid molecule comprising a first strand and a second strand;
- and
- a stem-loop oligonucleotide comprising:
- an inverted repeat;
- a loop;
- a non-replicable base or sequence, or at least one deoxy-uridine;
- and
- a 5′ end that lacks a phosphate;
- wherein the stem-loop oligonucleotide is attached to the first strand of the double stranded nucleic acid molecule and the 5′ end of the stem-loop oligonucleotide is separated from the second strand of the double stranded nucleic acid molecule.
A composition comprising a plurality of oligonucleotide-attached nucleic acid molecules each comprising:
- a double stranded nucleic acid molecule;
- and
- a stem-loop oligonucleotide comprising an inverted repeat and a loop, wherein one strand of the stem-loop oligonucleotide is attached to one strand of the double stranded nucleic acid molecule and wherein the stem loop oligonucleotide further comprises at least one deoxy-uridine.
Dependencies are inferred from the claim text.