Methods of producing nucleic acids using oligonucleotides modified by a stimulus
AssigneeTakara Bio USA, Inc.
InventorKazuo Tori
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Abstract
Provided are methods of producing product nucleic acids involving the use of oligonucleotides that are modified by the application of a stimulus. Aspects of such methods may include producing product nucleic acids using de-activatable oligonucleotides that are deactivated by a de-activating stimulus, as well as methods that may include producing product nucleic acids using activatable oligonucleotides that are activated by an activating stimulus and de-activatable oligonucleotides that are deactivated by a de-activating stimulus. Also provided are kits, compositions and devices that include de-activatable oligonucleotides or activatable and de-activatable oligonucleotides, e.g., for use in performing the methods as described herein.
Claims14
A method comprising:
- elongating a primer oligonucleotide via a template nucleic acid-mediated primer extension reaction, wherein the elongating comprises a template switching reaction comprising a de-activatable template switching oligonucleotide (TSO) to produce an extended nucleic acid that comprises a sequence complementary to the TSO;
- and
- deactivating the de-activatable oligonucleotide, wherein the de-activatable oligonucleotide comprises a cleavable-base.
A method comprising:
- elongating a primer oligonucleotide via a template nucleic acid-mediated primer extension reaction, wherein the elongating comprises a template switching reaction comprising a de-activatable template switching oligonucleotide (TSO) to produce an extended nucleic acid that comprises a sequence complementary to the TSO;
- deactivating the de-activatable oligonucleotide;
- amplifying the extended nucleic acid to produce an amplification product;
- and
- sequencing an amplification product of the amplification reaction.
A method comprising:
- elongating a primer oligonucleotide via a template nucleic acid-mediated primer extension reaction, wherein the elongating comprises a template switching reaction comprising a de-activatable template switching oligonucleotide (TSO) to produce an extended nucleic acid that comprises a sequence complementary to the TSO;
- and
- deactivating the de-activatable oligonucleotide,
- wherein the method is performed in a single reaction vessel and the method further comprises lysing a single cell in the single reaction vessel.
Dependencies are inferred from the claim text.