| CPC G01N 33/6818 (2013.01) [C12Q 1/68 (2013.01)] | 29 Claims |
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1. A method of sequencing a peptide at single amino acid resolution, comprising:
(a) providing said peptide and a linker, wherein said linker comprises a polymerizable molecule, and wherein said linker is capable of coupling to an amino acid of said peptide;
(b) coupling said linker to said amino acid of said peptide;
(c) coupling said linker to a capture moiety;
(d) cleaving said amino acid from said peptide to yield an amino acid-linker-capture moiety (AALC) complex;
(e) providing an additional linker, wherein said additional linker is capable of coupling to an additional amino acid of said peptide;
(f) coupling said additional linker to said additional amino acid, thereby generating an additional amino acid-linker complex; and
(g) coupling said additional amino acid-linker complex to said AALC complex, thereby generating a stacked AALC complex;
(h) contacting said stacked AALC complex with a plurality of binding agents, wherein individual binding agents of said plurality of binding agents have different specificity to different amino acid types;
(i) translocating said stacked AALC complex contacted with said plurality of binding agents through a nanopore or a nanogap; and
(j) identifying an amino acid type of said amino acid or said additional amino acid.
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