Single molecule loading methods and compositions
First Claim
Patent Images
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;
providing the population of target molecules to be distributed;
adjusting the size of the confined reaction or observation regions by adding at least one sizing moiety to individual reaction or observation regions such that a selected number of target molecules will fit into the resulting size-adjusted regions, oradjusting the size of individual target molecules of the population by linking at least one 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; and
,loading the target molecules into the regions, whereby a selected number of target molecules can fit into each region, thereby distributing the population of target molecules into the plurality of size confined regions.
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Abstract
Methods, compositions and arrays for non-random loading of single analyte molecules into array structures are provided.
174 Citations
164 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:
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providing a structure comprising the plurality of size confined reaction or observation regions; providing the population of target molecules to be distributed; adjusting the size of the confined reaction or observation regions by adding at least one sizing moiety to individual reaction or observation regions such that a selected number of target molecules will fit into the resulting size-adjusted regions, or adjusting the size of individual target molecules of the population by linking at least one 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; and
,loading the target molecules into the regions, whereby a selected number of target molecules can fit into each region, thereby distributing the population of target molecules into the plurality of size confined regions. - View Dependent Claims (3, 4, 15, 20, 24, 25, 27)
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2. (canceled)
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41. 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 particles that comprise, bind or package a population of target molecules, 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; and
;delivering the population of particles into the plurality of wells, thereby distributing the population of target molecules into the plurality of wells of the small well array.
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42-62. -62. (canceled)
- 63. An analysis device comprising an array of analyte molecules, wherein the analyte molecules are arranged in the array by one or more phase determining feature such that a single molecule of the analyte is present in at least 40% of the analysis regions of the array.
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66-84. -84. (canceled)
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85. A method of producing a non-random distribution of single analyte molecules in analysis regions of an array, the method comprising:
selectively distributing the analyte molecules into the analysis regions, such that at least 38% of the regions are occupied by one analyte molecule, fewer than 5% of the analysis regions are occupied by more than one analyte molecule, and fewer than 62% of the analysis regions are occupied by fewer than one analyte molecule, thereby producing the non-random distribution of analyte molecules in the analysis regions of the array. - View Dependent Claims (86, 87, 88, 100, 103, 104, 105, 106, 123, 124, 125, 128, 130, 131, 132, 136, 155, 156, 157, 161, 162, 163)
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Specification