METHOD AND COMPOSITIONS FOR DETECTION AND ENUMERATION OF GENETIC VARIATIONS

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Many areas of biomedical research depend on the analysis of uncommon variations in individual genes or transcripts. Here we describe a method tha t can quantify such variation at a scale and ease heretofore unattainable. Eac h DNA molecule in a collection of such molecules is converted into a single particle to which thousands of copies of DNA identical in sequence to the original are bound. This population of beads then corresponds to a one-to-on e representation of the starting DNA molecules. Variation within the original population of DNA molecules can then be simply assessed by counting fluorescently-labeled particles via flow cytometry. Millions of individual D NA molecules can be assessed in this fashion with standard laboratory equipment . Moreover, specific variants can be isolated by flow sorting and employed for further experimentation. This approach can be used for the identification an d quantification of rare mutations as well as to study variations in gene sequences or transcripts in specific populations or tissues.