Methods of making nucleic acid molecules with reduced secondary structure
First Claim
1. A method of synthesizing an unstructured nucleic acid with reduced secondary structure relative to a nucleic acid of substantially identical nucleotide sequence having naturally occurring bases, the method comprising steps of:
- providing a template nucleic acid strand comprising at least two self-complementary sequence elements;
selecting a DNA polymerase for use in synthesizing a new unstructured nucleic acid strand complementary to the template nucleic acid strand so that the new unstructured strand contains at least two self-complementary sequence elements;
providing a collection of nucleotides that are substrates for the DNA polymerase, the collection being selected to allow for intermolecular base-pair interactions but not intermolecular base pair interactions; and
allowing extension by the DNA polymerase, so that the new unstructured nucleic acid strand is synthesized and the self-complementary sequence elements within the unstructured nucleic acid strand do not form intramolecular hybrids with one another but do form intermolecular hybrids with the template strand.
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Abstract
The present invention provides a system for generating nucleic acid molecules having reduced levels of secondary structure compared to nucleic acid molecules of the same nucleotide sequence containing only naturally-occurring bases. Such molecules are referred to herein as “unstructured nucleic acids” (UNAs). UNAs have reduced levels of secondary structure because of their reduced ability to form intramolecular hydrogen bond base pairs between regions of substantially complementary sequence. Preferred UNAs, however, retain the ability to form intermolecular hydrogen bond base pairs with other nucleic acid molecules.
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Citations
7 Claims
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1. A method of synthesizing an unstructured nucleic acid with reduced secondary structure relative to a nucleic acid of substantially identical nucleotide sequence having naturally occurring bases, the method comprising steps of:
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providing a template nucleic acid strand comprising at least two self-complementary sequence elements;
selecting a DNA polymerase for use in synthesizing a new unstructured nucleic acid strand complementary to the template nucleic acid strand so that the new unstructured strand contains at least two self-complementary sequence elements;
providing a collection of nucleotides that are substrates for the DNA polymerase, the collection being selected to allow for intermolecular base-pair interactions but not intermolecular base pair interactions; and
allowing extension by the DNA polymerase, so that the new unstructured nucleic acid strand is synthesized and the self-complementary sequence elements within the unstructured nucleic acid strand do not form intramolecular hybrids with one another but do form intermolecular hybrids with the template strand. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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Specification