Compositions and kits for molecular counting
AssigneeBecton, Dickinson and Company
InventorsGlenn K. Fu, Stephen P.A. Fodor, Julie Wilhelmy
Life of this patent
Estimating the term…
Abstract
Methods, kits and systems are disclosed for analyzing one or more molecules in a sample. Analyzing the one or more molecules may comprise quantitation of the one or more molecules. Individual molecules may quantitated by PCR, arrays, beads, emulsions, droplets, or sequencing. Quantitation of individual molecules may further comprise stochastic labeling of the one or more molecules with a plurality of oligonucleotide tags to produce one or more stochastically labeled molecules. The methods may further comprise amplifying, sequencing, detecting, and/or quantifying the stochastically labeled molecules. The molecules may be DNA, RNA and/or proteins.
Claims21
-163. (canceled)
A composition for counting the number of occurrences of one or more mRNA target molecules, comprising:
- a plurality of first labeled double-stranded DNA molecules corresponding to copies of a first mRNA target molecule, wherein each of the plurality of first labeled double-stranded DNA molecules comprises:
- a target specific sequence comprising an oligo dT sequence,
- a unique identifier sequence,
- a universal PCR primer sequence, and
- a copy of the first mRNA target molecule, a complement thereof, a reverse complement thereof, or a portion of any of the preceding,
- wherein each of the plurality of first labeled double-stranded DNA molecules is produced by nested PCR using the universal PCR primer sequence
- wherein the unique identifier sequence uniquely identifies occurrences of the mRNA target molecule, and
- wherein the number of different unique identifier sequences of the plurality of first labeled double-stranded DNA molecules indicates the number of occurrences of the first mRNA target molecule.
A kit for counting the number of occurrences of one or more mRNA target molecules, comprising:
- a plurality of oligonucleotide tags, wherein each oligonucleotide tag of the plurality of oligonucleotide tags comprises:
- a target specific sequence comprising an oligo dT sequence,
- a unique identifier sequence,
- a universal PCR primer sequence, and
- wherein at least 1000 of the plurality of oligonucleotide tags comprise different unique identifier sequence;
- a universal primer capable of binding to the oligonucleotide tag or the universal primer binding site of the oligonucleotide tag;
- and
- an enzyme.
Dependencies are inferred from the claim text.