DIGITAL COUNTING OF INDIVIDUAL MOLECULES BY STOCHASTIC ATTACHMENT OF DIVERSE LABELS

摘要

Compositions, methods and kits are disclosed for high-sensitivity single molecule digital counting by the stochastic labeling of a collection of identical molecules by attachment of a diverse set of labels. Each copy of a molecule randomly chooses from a non-depleting reservoir of diverse labels. Detection may be by a variety of methods including hybridization based or sequencing. Molecules that would otherwise be identical in information content can be labeled to create a separately detectable product that is unique or approximately unique in a collection. This stochastic transformation relaxes the problem of counting molecules from one of locating and identifying identical molecules to a series of binary digital questions detecting whether preprogrammed labels are present. The methods may be used, for example, to estimate the number of separate molecules of a given type or types within a sample.

主权项

1. A composition comprising a plurality of beads, wherein individual beads comprise a plurality of attached oligonucleotides, and wherein the plurality of attached oligonucleotides comprise a set, m, of variable target label sequences capable of attaching to or hybridizing to a plurality, n, of target molecules in a stochastic manner, wherein m is greater than n, and wherein the plurality of attached oligonucleotides are capable of being used for stochastic labeling of one or more different types of target molecules.