Single molecule loading methods and compositions
First Claim
1. A method of distributing a population of target molecules into a plurality of size confined reaction or observation regions, the method comprising:
- providing a structure comprising the plurality of size confined reaction or observation regions, wherein such regions comprise wells;
providing the population of target molecules to be distributed;
linking at least one nucleic acid sizing moiety to individual target molecules, wherein the sizing moieties are of sufficient size, relative to the size-confined reaction or observation regions, such that only a single sizing moiety will fit into each size confined region, thereby preventing the entry of an additional sizing moiety; and
,loading the target molecules into the regions such that the size of the confined regions results in a single target molecule distributed into each region, thereby distributing the population of target molecules into the plurality of size confined regions.
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Abstract
Methods for non-random loading of single analyte molecules into array structures are provided. The methods allow for distribution of a population of target molecules into a plurality of size confined regions such as wells. Sizing moieties are linked to individual target molecules. The sizing moieties are of sufficient size, relative to the size-confined reaction or observation regions, such that only a selected number of sizing moieties will fit into the size confined regions. The confined regions and the sizing moieties or target molecules comprise a selected charge that allow for controlling the loading of the sizing moities.
34 Citations
15 Claims
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1. A method of distributing a population of target molecules into a plurality of size confined reaction or observation regions, the method comprising:
providing a structure comprising the plurality of size confined reaction or observation regions, wherein such regions comprise wells; providing the population of target molecules to be distributed; linking at least one nucleic acid sizing moiety to individual target molecules, wherein the sizing moieties are of sufficient size, relative to the size-confined reaction or observation regions, such that only a single sizing moiety will fit into each size confined region, thereby preventing the entry of an additional sizing moiety; and
,loading the target molecules into the regions such that the size of the confined regions results in a single target molecule distributed into each region, thereby distributing the population of target molecules into the plurality of size confined regions. - View Dependent Claims (2, 3, 4, 5, 7, 8, 9, 10, 11, 12)
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6. A method of distributing a population of target molecules into a plurality of wells in a small well array, the method comprising:
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providing the small well array, which array comprises the plurality of wells, which wells individually comprise a reaction or observation region; providing a population of nucleic acid particles that comprise, bind, or package a single target molecule, wherein the plurality of wells individually are configured to receive a single particle from the population of particles into the reaction or observation region of the well wherein the nucleic acid particles prevents the entry of additional nucleic acid particles; and
;delivering the population of particles into the plurality of wells, thereby distributing a single target molecule into the plurality of wells of the small well array.
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13. A method of distributing a population of target molecules into a plurality of size confined reaction or observation regions, the method comprising:
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providing a structure comprising the plurality of size confined reaction or observation regions, wherein such regions comprise wells; providing the population of target molecules to be distributed; linking at least one nucleic acid sizing moiety to individual target molecules, wherein the sizing moieties are of sufficient size, relative to the size-confined reaction or observation regions, so that only a selected number of sizing moieties will fit into the size confined regions, wherein the nucleic acid sizing moiety comprises DNA origami; and
,loading the target molecules into the regions such that the size of the confined regions results in a single target molecule distributed into each region, thereby distributing the population of target molecules into the plurality of size confined regions. - View Dependent Claims (14, 15)
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Specification