Methods and systems for nanopore sequencing of modified amino acids

A method for measuring a signal of a stacked plurality of modified amino acids, comprising:

• (a) providing said stacked plurality of modified amino acids, wherein said stacked plurality of modified amino acids comprises a plurality of modified amino acids comprising a plurality of nucleic acid molecules coupled thereto, wherein said plurality of nucleic acid molecules is coupled to one another in a linear fashion to produce a linear nucleic acid molecule, wherein said linear nucleic acid molecule comprises a double stranded priming region;
• (b) contacting said stacked plurality of modified amino acids with a plurality of nucleotides and a polymerizing enzyme under conditions sufficient to perform a primer extension reaction of said double stranded priming region, thereby generating a partially polymerized stacked plurality of modified amino acids;
• (c) translocating, in a first direction, said partially polymerized stacked plurality of modified amino acids or a portion thereof through a nanopore;
• (d) translocating, in a second direction, said partially polymerized stacked plurality of modified amino acids or a portion thereof through said nanopore;
• and
• (e) repeating (b)-(d) on said partially polymerized stacked plurality of modified amino acids;
• wherein, during (d), said signal is measured from or adjacent to said nanopore, thereby generating a measured signal corresponding to an identity of a modified amino acid of said plurality of modified amino acids.