Back to results
US 12,668,833 B2Granted
Methods, systems, and compositions for counting nucleic acid molecules
AssigneeEnumera Molecular, Inc.
InventorsMatthew Sekedat, Jeffrey Buis, Ronald David Beaubien, Sharat Singh, Jeff Perry
Life of this patent
Estimating the term…
Abstract
Compositions and methods, systems, and kits for detecting and quantifying variations in numbers of molecules, particularly variations in gene dosage, e.g., due to gene duplication, or to variations from the normal euploid complement of chromosomes, e.g., trisomy of one or more chromosomes that are normally found in diploid pairs, without digital sequencing.
Claims20
A method, comprising:
- a) forming at least one complex comprising an oligonucleotide primer hybridized to a circularized nucleic acid, wherein the primer is bound to a surface on a solid support;
- and
- b) exposing the at least one complex to a reaction mixture comprising a DNA polymerase, one or more deoxynucleoside triphosphates (dNTPs), and at least 12% polyethylene glycol (PEG), wherein the PEG has an average molecular weight between 400 and 800, under conditions wherein the primer in the complex is extended on the circularized nucleic acid by the DNA polymerase to form an extended primer.
A composition comprising:
- a) a plurality of complexes bound to a silanized surface of a solid support, each complex comprising an oligonucleotide primer hybridized to a circularized nucleic acid, wherein primers in the complexes are covalently linked to the silanized surface of the solid support in a dispersal;
- and
- b) a reaction mixture in contact with the plurality of complexes, the reaction mixture comprising a DNA polymerase, one or more dNTPs, and at least 12% PEG, wherein the PEG has an average molecular weight between 400 and 800.
Dependencies are inferred from the claim text.