Methods for simultaneous generation of functional ligands
First Claim
1. A method for simultaneously selecting a plurality of aptamers for multiple different targets comprising:
- contacting a library containing aptamers comprising a diversity of nucleic acids each having a randomized region and a constant region, with a plurality of different targets simultaneously in a single reaction volume, each of said targets being fixed at a different predetermined location on an array;
partitioning members of said library that do not bind to said plurality of different targets;
marking the members of the library that bind to the plurality of different targets with identifiers, each of said identifiers being indicative of the particular target;
applying a different identifier to each of said different predetermined locations on said array, said identifier comprising an single-stranded DNA oligonucleotide having a variable region which identifies one of said predetermined locations on said array, said variable region being flanked by a first constant region having a sequence for hybridizing to said constant region of said library, and a second constant region having a primer binding site;
identifying the binding members of the library and correlating them to the targets via said identifiers; and
performing a nucleic acid amplification reaction on each of said hybridized identifiers and said aptamers to generate an amplification product;
wherein said amplification product correlates the sequence of one of said aptamers to one of said targets to which said aptamer binds.
2 Assignments
0 Petitions
Accused Products
Abstract
The present invention relates to methods for generating functional biomolecules. In one exemplary aspect of the invention, generation of functional biomolecules may be performed against multiple targets simultaneously within a single system. In general, a plurality of targets may be disposed within in a single reaction volume and a library of biomolecules, such as a nucleic acid library, may be applied to the reaction volume. The members of the library that do not bind to any of the plurality of targets under given conditions may then be partitioned. The remaining members of the library may then be marked and/or tagged, such as to identify the particular target or targets to which the member of the library binds. The binding members of the library may then be isolated and, by virtue of the marking or tagging, be matched to a particular target or targets.
-
Citations
15 Claims
-
1. A method for simultaneously selecting a plurality of aptamers for multiple different targets comprising:
-
contacting a library containing aptamers comprising a diversity of nucleic acids each having a randomized region and a constant region, with a plurality of different targets simultaneously in a single reaction volume, each of said targets being fixed at a different predetermined location on an array; partitioning members of said library that do not bind to said plurality of different targets; marking the members of the library that bind to the plurality of different targets with identifiers, each of said identifiers being indicative of the particular target; applying a different identifier to each of said different predetermined locations on said array, said identifier comprising an single-stranded DNA oligonucleotide having a variable region which identifies one of said predetermined locations on said array, said variable region being flanked by a first constant region having a sequence for hybridizing to said constant region of said library, and a second constant region having a primer binding site; identifying the binding members of the library and correlating them to the targets via said identifiers; and performing a nucleic acid amplification reaction on each of said hybridized identifiers and said aptamers to generate an amplification product; wherein said amplification product correlates the sequence of one of said aptamers to one of said targets to which said aptamer binds. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
-
Specification