Methods and apparatus for analyzing polynucleotide sequences
First Claim
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1. A method of analyzing a target polynucleotide comprising:
- (a) pretreating the surface of a substrate with a polyelectrolyte multilayer (PEM) to create surface chemistry that facilitates polynucleotide attachment and sequence analysis;
(b) providing a primed target polynucleotide attached to a surface of a substrate;
(c) providing a labeled first nucleotides to the attached target polynucleotide under conditions whereby the labeled first nucleotide attaches to the primer, if a complementary nucleotide is present to serve as template in the target polynucleotide;
(d) determining presence or absence of a signal, the presence of a signal indicating that the labeled first nucleotide was incorporated into the primer, and hence the identity of the complementary base that served as a template in the target polynucleotide;
(e) repeating steps (c)-(d) with a labeled further nucleotide, the same or different from the first labeled nucleotide, whereby the labeled further nucleotide attaches to the primer or a nucleotide previously incorporated into the primer; and
(f) repeating step (e) until identities of the bases in a portion or all of the target polynucleotide are determined.
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Abstract
Methods for high speed, high throughput analysis of polynucleotide sequences, and apparatuses with which to carry out the methods are provided in the invention.
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Citations
1 Claim
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1. A method of analyzing a target polynucleotide comprising:
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(a) pretreating the surface of a substrate with a polyelectrolyte multilayer (PEM) to create surface chemistry that facilitates polynucleotide attachment and sequence analysis;
(b) providing a primed target polynucleotide attached to a surface of a substrate;
(c) providing a labeled first nucleotides to the attached target polynucleotide under conditions whereby the labeled first nucleotide attaches to the primer, if a complementary nucleotide is present to serve as template in the target polynucleotide;
(d) determining presence or absence of a signal, the presence of a signal indicating that the labeled first nucleotide was incorporated into the primer, and hence the identity of the complementary base that served as a template in the target polynucleotide;
(e) repeating steps (c)-(d) with a labeled further nucleotide, the same or different from the first labeled nucleotide, whereby the labeled further nucleotide attaches to the primer or a nucleotide previously incorporated into the primer; and
(f) repeating step (e) until identities of the bases in a portion or all of the target polynucleotide are determined.
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Specification