Varietal counting of nucleic acids for obtaining genomic copy number information

摘要

A method for obtaining from genomic material genomic copy number information unaffected by amplification distortion, comprising obtaining segments of the genomic material, tagging the segments with substantially unique tags to generate tagged nucleic acid molecules, such that each tagged nucleic acid molecule comprises one segment of the genomic material and a tag, subjecting the tagged nucleic acid molecules to polymerase chain reaction (PCR) amplification, generating tag associated sequence reads by sequencing the product of the PCR reaction, assigning each tagged nucleic acid molecule to a location on a genome associated with the genomic material by mapping the subsequence of each tag associated sequence read corresponding to a segment of the genomic material to a location on the genome, and counting the number of tagged nucleic acid molecules assigned to the same location on the genome having a different tag, thereby obtaining genomic copy number information unaffected by amplification distortion.

主权项

1. A method for obtaining genomic copy number information from genomic nucleic acids, comprising:
a) obtaining segments of the genomic nucleic acids; b) tagging the segments with substantially unique tags, thereby generating tagged nucleic acid molecules, such that each of the tagged nucleic acid molecules comprises one segment of the segments of the genomic nucleic acids from step (a) and a substantially unique tag having a unique nucleotide sequence of the substantially unique tags, so as to permit one substantially unique tag having a unique nucleotide sequence of the substantially unique tags to differentiate from another tag having a unique nucleotide sequence of the substantially unique tags; c) subjecting the tagged nucleic acid molecules to a polymerase chain reaction (PCR), thereby generating amplified tagged nucleic acid molecules; d) generating tag associated sequence reads by sequencing the amplified tagged nucleic acid molecules of step (c); e) assigning each of the amplified tagged nucleic acid molecules to a location on a genome associated with the genomic nucleic acids by mapping the subsequence of each tag associated sequence read of said tag associated sequence reads of step (d) corresponding to one segment of the segments of the genomic nucleic acids to a location on the genome; and f) in the amplified tagged nucleic acid molecules, counting the number of the amplified tagged nucleic acid molecules having a different substantially unique tag that have been assigned to the same location on the genome, thereby obtaining counts of the amplified tagged nucleic acid molecules having a different substantially unique tag that have been assigned to the same location of the genome and the genomic copy number information from the genomic nucleic acids, wherein the different substantially unique tag is from the substantially unique tags, and wherein the genome has multiple locations.