Polymer sequencing using selectively labeled monomers and data integration
First Claim
1. A method of sequencing a polymer comprising:
- a) dividing a polymer sample into a number of polymer subsamples equal to a number of different monomer types comprising the polymer sample, wherein only one of the monomer types in each polymer subsample is partially labeled such that an average time between two adjacent labeled monomers is significantly larger than an average time between two adjacent monomers of the same type in the polymer subsample before labelling;
b) sequentially separating each monomer from the polymer subsample;
c) detecting the labels of each separated labeled monomer as a function of time;
d) constructing a time map for each monomer type in each polymer subsample; and
e) assembling the time maps into a polymer sequence.
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Abstract
Methods and apparatuses for sequencing single polymer molecules, such as a nucleic acid strand, are discussed. A discussed method comprises dividing a polymer sample into a number of polymer subsamples equal to the number of different monomer types and partially labeling only one of the monomer types in each polymer subsample. The method may further comprise placing a subsample into a reaction chamber, sequentially separating each monomer from the polymer subsample, and detecting the labels of each separated labeled monomer as a function of time. The time between each labeled monomer may be used to construct a monomer-time map for each polymer sub-sample using overlapping data analysis and frequency analysis. Time maps may then be assembled/aligned into a polymer sequence from the monomer-time maps of each of the polymer subsamples using non-overlapping data analysis.
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Citations
29 Claims
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1. A method of sequencing a polymer comprising:
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a) dividing a polymer sample into a number of polymer subsamples equal to a number of different monomer types comprising the polymer sample, wherein only one of the monomer types in each polymer subsample is partially labeled such that an average time between two adjacent labeled monomers is significantly larger than an average time between two adjacent monomers of the same type in the polymer subsample before labelling;
b) sequentially separating each monomer from the polymer subsample;
c) detecting the labels of each separated labeled monomer as a function of time;
d) constructing a time map for each monomer type in each polymer subsample; and
e) assembling the time maps into a polymer sequence. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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13. A method of sequencing a polymer comprising:
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a) dividing a polymer sample into a number of polymer subsamples equal to a number of different monomer types comprising the polymer sample, wherein only one of the monomer types in each polymer subsample is partially labeled such that an average time between two adjacent labeled monomers is significantly larger than an average time between two adjacent monomers of the same type in the polymer subsample before labelling;
b) moving the intact partially labeled polymer across a detector;
c) measuring a time between the partially labeled monomers;
d) constructing a time map for each detected labeled monomer for each partially labeled polymer strand; and
e) assembling the time maps into a sequence for the polymer. - View Dependent Claims (14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
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25. An apparatus comprising:
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a) a chamber for cleaving a partially-labeled polymer sample into individual monomers;
b) a means for transporting the individual monomers across a detector. - View Dependent Claims (26, 27, 28, 29)
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Specification