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US 12,037,707 B2Granted
Methods of making and using combinatorial barcoded nucleic acid libraries having defined variation
AssigneesMassachusetts Eye and Ear Infirmary, The Schepens Eye Research Institute, Inc.
InventorsLuk H. Vandenberghe, Eric Zinn, Heikki Turunen
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Abstract
This disclosure describes compositions, methods, and systems for constructing defined variation in a contiguous functional genetic unit in association with a unique sequence identifier (“a barcode”) in a combinatorial manner.
Claims19
A method of generating a combinatorial library, the method comprising:
- providing a first acceptor sequence comprising a first portion of an acceptor target sequence and a first portion of a corresponding acceptor barcode sequence;
- providing a first donor sequence comprising a first portion of a donor target sequence and a first portion of a corresponding donor barcode sequence;
- cleaving the first acceptor sequence between the first portion of the acceptor target sequence and the first portion of the corresponding acceptor barcode sequence;
- and
- ligating the first donor sequence into the cleaved first acceptor sequence,
- thereby producing a second acceptor sequence comprising a second portion of an acceptor target sequence and a second portion of a corresponding acceptor barcode sequence.
3 more elements
Dependencies are inferred from the claim text.