High resolution linear analysis of polymers
First Claim
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1. A method for analyzing a polymer comprising:
- providing the polymer having first and second unit specific markers, the first unit specific marker including a first label and the second unit specific marker including a second label, wherein the first and second unit specific markers are spaced apart on the polymer by a separation distance;
providing a detection zone adapted to detect emission signals, the detection zone characterized by a zone distance;
moving the polymer through the detection zone at a velocity;
detecting signals emitted from the label of the first unit specific marker before and after a first timing event as the first unit specific marker passes through the detection zone, to define a first emission signal;
calculating a proportion of the first emission signal that corresponds to a distance of the detection zone traversed by the label of the first unit specific marker at the first timing event;
detecting signals emitted from the label of the second unit specific marker before and after a second timing event as the second unit specific marker passes through the detection zone, to define a second emission signal;
calculating a proportion of the second emission signal that corresponds to a distance of the detection zone traversed by the label of the second unit specific marker at the second timing event;
determining the separation distance at a precision greater than the zone distance by comparing the proportion of the first emission signal and the proportion of the second emission signal; and
outputting an indication of the separation distance to a user.
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Abstract
The invention provides methods and systems for improved spatial resolution of signal detection, particularly as applied to the analysis of polymers such as biological polymers. Some of the methods and systems comprise differentially tagging polymers in order to increase resolution. Some of the methods and systems comprise techniques for improving the precision of separation distance measurements, without necessarily requiring improvements in the known detection resolution of prior art systems.
24 Citations
35 Claims
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1. A method for analyzing a polymer comprising:
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providing the polymer having first and second unit specific markers, the first unit specific marker including a first label and the second unit specific marker including a second label, wherein the first and second unit specific markers are spaced apart on the polymer by a separation distance; providing a detection zone adapted to detect emission signals, the detection zone characterized by a zone distance; moving the polymer through the detection zone at a velocity; detecting signals emitted from the label of the first unit specific marker before and after a first timing event as the first unit specific marker passes through the detection zone, to define a first emission signal; calculating a proportion of the first emission signal that corresponds to a distance of the detection zone traversed by the label of the first unit specific marker at the first timing event; detecting signals emitted from the label of the second unit specific marker before and after a second timing event as the second unit specific marker passes through the detection zone, to define a second emission signal; calculating a proportion of the second emission signal that corresponds to a distance of the detection zone traversed by the label of the second unit specific marker at the second timing event; determining the separation distance at a precision greater than the zone distance by comparing the proportion of the first emission signal and the proportion of the second emission signal; and outputting an indication of the separation distance to a user. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34)
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35. A method comprising:
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moving through a detection zone a polymer bound to a first labeled unit specific marker and a second labeled unit specific marker, wherein the first and second unit specific markers are spaced apart by a separation distance, detecting signals that define a first emission signal and are emitted from the first labeled unit specific marker, before and after a first timing event, as the first labeled unit specific marker pass through the detection zone, detecting signals that define a second emission signal and are emitted from the second labeled unit specific marker, before and after a second timing event, as the second labeled unit specific marker passes through the detection zone, calculating a proportion of the first emission signal that corresponds to a distance of the detection zone traversed by the first labeled unit specific marker at the first timing event, calculating a proportion of the second emission signal that corresponds to a distance of the detection zone traversed by the second labeled unit specific marker at the second timing event, determining the separation distance at a precision greater than the zone distance by comparing the proportion of the first emission signal and the proportion of the second emission signal; and outputting an indication of the separation distance to a user.
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Specification