Digital analyte analysis

摘要

The invention generally relates to droplet based digital PCR and methods for analyzing a target nucleic acid using the same. In certain embodiments, a method for determining the nucleic acid make-up of a sample is provided.

主权项

1. A method for determining a number of nucleic acids that comprise both first and second locations, the method comprising:
partitioning a sample comprising a plurality of nucleic acids of different lengths into partitioned portions, wherein a plurality of the partitioned portions each contain a single nucleic acid; introducing first and second primer pairs and first and second detectably labeled probes to each of the partitioned portions, wherein the first and second primer pairs are specific for first and second locations on the nucleic acid, the first and second locations being spaced apart from each other, and wherein the first probe hybridizes to the first location and the second probe hybridizes to the second location on the nucleic acid; amplifying the nucleic acids in the partitioned portions; detecting amplicons in the partitioned portions, the presence of signal from both probes indicating the presence of a nucleic acid that is contiguous between the first and second locations on the nucleic acid; and determining the number of nucleic acids that comprise both the first and second locations based upon results of the detecting step.