Self-assembled single molecule arrays and uses thereof

A method of making a probe array, the method comprising the steps of:

• generating a plurality of polynucleotide molecules each comprising a concatemer of a probe sequence and an adaptor oligonucleotide;
• disposing the plurality of polynucleotide molecules onto a support having a surface with capture oligonucleotides attached thereto so that the polynucleotide molecules are fixed to the surface by one or more complexes formed between capture oligonucleotides and adaptor oligonucleotides and so that the polynucleotide molecules are randomly distributed on the surface at a density such that a majority of the polynucleotide molecules have a nearest neighbor distance of at least fifty run, thereby forming the array of polynucleotide molecules;
• and
• identifying the probe sequence of each polynucleotide molecule on the surface to form the probe array.

A method of measuring amounts of a plurality of different target polynucleotides in a sample, the method comprising the steps of:

• providing a pool of nucleic acid fragments derived from the sample containing the plurality of different target polynucleotides;
• hybridizing the pool of nucleic acid fragments to a random array of probe concatemers fixed to a planar surface having an array of optically resolvable discrete spaced apart regions, wherein each discrete spaced apart region has an area of less than I μm2 and wherein substantially all such regions have at most one of said probe concatemers attached, each probe concatemer comprising multiple copies of a fragment of a target polynucleotide of the plurality, or a complement thereof, where the fragment or complement thereof is identified by a portion of its sequence;
• and
• quantifying the hybridization of said nucleic acid fragments to the random array, wherein such quantification is proportional to the amounts of the plurality of different polynucleotides in the sample.

A method of measuring amounts of a plurality of different target polynucleotides in a sample, the method comprising the steps of:

• providing a pool of nucleic acid fragments derived from the sample containing the plurality of different target polynucleotides;
• hybridizing the pool of nucleic acid fragments to a random array of probe concatemers fixed to a surface at a density such that at least a majority of the probe concatemers are optically resolvable, each probe concatemer comprising multiple copies of a fragment of a target polynucleotide of the plurality or a complement thereof, where the fragment or complement thereof is identified by a portion of its sequence;
• and
• quantifying the hybridization of said nucleic acid fragments to the random array, wherein such quantification is proportional to the amounts of the plurality of different polynucleotides in the sample.