Methods for retrieval of sequence-verified DNA constructs
AssigneeUniversity of Washington through its Center for Commercialization
InventorsJay Shendure, Jerrod Schwartz, Jacob Kitzman, Rupali Patwardhan, Joseph Hiatt
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Abstract
In some embodiments, methods of recovering a sequence-verified target nucleic acid are provided. In some embodiments, such methods may include tagging each member of a nucleic acid library with a set of adaptor sequences; sequencing the tagged members of the nucleic acid library; and recovering the sequence-verified target nucleic acid from the tagged and sequenced members of the nucleic acid library using a dial-out selection method. In certain embodiments, the members of the nucleic acid library may be tagged with a second set of adaptor sequences.
Claims16
A method of recovering a sequence-verified target nucleic acid from a sequenced nucleic acid library comprising
- tagging each member of a nucleic acid library with at least one identification tag having a degenerate sequence;
- sequencing the tagged members of the nucleic acid library;
- and
- selectively recovering only the sequence-verified target nucleic acid from the tagged and sequenced members of the nucleic acid library by targeting the at least one identification tag that is unique to the sequence-verified target nucleic acid using a clone-free dial-out selection method and amplifying the sequence-verified target nucleic acids.
A method of recovering a sequence-verified target nucleic acid variant comprising:
- tagging each member of a mutagenesis library with at least one identification tag having a degenerate sequence;
- sequencing the tagged members of the mutagenesis library;
- and
- selectively recovering only the sequence-verified target nucleic acid variant from the tagged and sequenced members of the mutagenesis library by targeting the at least one identification tag that is unique to the sequence-verified target nucleic acid variant using a clone-free dial-out selection method and amplifying the sequence-verified target nucleic acids.
A method of synthesizing a DNA construct comprising:
- tagging each member of a nucleic acid library with at least one identification tag having a degenerate sequence;
- sequencing the tagged members of the nucleic acid library;
- and
- selectively recovering only two or more sequence-verified target nucleic acids from the tagged and sequenced members of the nucleic acid library by targeting the at least one identification tag that is unique to the two or more sequence-verified target nucleic acids using a clone-free dial-out selection method and amplifying the two or more sequence-verified target nucleic acids;
- and
- assembling the DNA construct using the two or more sequence-verified target nucleic acids.
Dependencies are inferred from the claim text.