Population Genetics Technologies Ltd.

Agilent Technologies, Inc.

Population Genetics Technologies Litd.

James Casbon

Robert Osborne

Andreas Claas

Sydney Brenner

Conrad Lichtenstein

Gi Mikawa

Esther Musgrave-Brown

Described herein is a method for adding a counter sequence to the individual polynucleotide molecules of an initial nucleic acid sample. After addition of the counter sequence, the sample may be amplified and the number of initial target molecules in the sample can be estimated by counting the number of counter sequences associated with the amplified target molecules.

Method for attaching a counter sequence to a nucleic acid sample

Aspects of the present invention include methods and compositions for determining the number of individual polynucleotide molecules originating from the same genomic region of the same original sample that have been sequenced in a particular sequence analysis configuration or process. In these aspects of the invention, a degenerate base region (DBR) is attached to the starting polynucleotide molecules that are subsequently sequenced (e.g., after certain process steps are performed, e.g., amplification and/or enrichment). The number of different DBR sequences present in a sequencing run can be used to determine/estimate the number of different starting polynucleotides that have been sequenced. DBRs can be used to enhance numerous different nucleic acid sequence analysis applications, including allowing higher confidence allele call determinations in genotyping applications.

Increased confidence of allele calls with molecular counting

Method of tagging using a split DBR

Method of adding a DBR by primer extension

Method for preparing a counter-tagged population of nucleic acid molecules

Method for accurately counting starting molecules

Increasing confidence of allele calls with molecular counting

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Hinxton, United Kingdom

James Casbon